MAX Enteric Viral Panel-NR - BD

 
MAX Enteric Viral Panel-NR - BD
MAX™ Enteric Viral Panel-NR
                                                                                                                         443987
For In Vitro Diagnostic Use                                                                                              P0230(04)
For use with the BD MAX™ System                                                                                            2019-05
                                                                                                                            English

English: pages 1 – 16             Español: páginas 17 – 36

INTENDED USE
The BD MAX™ Enteric Viral Panel-NR performed on the BD MAX System, is an automated in vitro diagnostic test for the direct
qualitative detection and differentiation of enteric viral pathogens. The BD MAX Enteric Viral Panel-NR detects nucleic acids from
•    Norovirus GI & GII
•    Rotavirus A
Testing is performed on unpreserved soft to diarrheal or Cary-Blair preserved stool specimens from symptomatic patients with
suspected acute gastroenteritis, enteritis or colitis. The test is performed directly on the specimen, utilizing real-time polymerase
chain reaction (PCR) for the amplification of relevant gene target DNA/RNA. The test utilizes fluorogenic gene-specific hybridization
probes for the detection of the amplified DNA.
This test is intended for use, in conjunction with clinical presentation, laboratory findings, and epidemiological information, as an
aid in the differential diagnosis of Norovirus GI & GII, and Rotavirus A, infections. Results of this test should not be used as the
sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with other
organisms that are not detected by this test, and may not be the sole or definitive cause of patient illness. Negative results in the
setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this test or
non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn’s disease.
SUMMARY AND EXPLANATION OF THE PROCEDURE
Organisms that cause enteric diseases represent a significant cause of morbidity and mortality worldwide. Enteric pathogens
enter the body through the gastrointestinal tract and typically are spread via contaminated food and water or contact with vomit or
feces. The Centers for Disease Control and Prevention (CDC) estimates that each year there are 48 million cases of foodborne
illness in the United States resulting in 128,000 hospitalizations and 3,000 deaths.1 In the developing world, these illnesses
cause approximately 2 million deaths annually in young children.2 Each of the causative agents may result in slightly different
symptomology, including abdominal cramps or pain, loss of appetite, nausea or vomiting; however, all result in diarrhea.3 Repeated
bouts of diarrhea and persistent diarrheal disease disrupt intestinal function and absorption, potentially leading to childhood
malnutrition and growth retardation.4
The BD MAX Enteric Viral Panel-NR performed on the BD MAX System, can provide results for the BD MAX Enteric Viral Panel-
NR targets for 24 specimens in approximately 3 hours. The BD MAX Enteric Viral Panel-NR simultaneously detects the pathogens
responsible for gastroenteritis due to Norovirus GI & GII, and Rotavirus A. The assay includes an internal Sample Processing
Control. The BD MAX Enteric Viral Panel-NR automates the testing process and minimizes operator intervention from the time the
sample is placed onto the BD MAX System until results are available.
A soft to diarrheal stool specimen is collected and transported to the laboratory, homogenized and looped into a BD MAX Enteric
Viral Panel Sample Buffer Tube. The Sample Buffer Tube is placed into the BD MAX System and the following automated
procedures occur: the viral capsids are lysed, DNA/RNA is extracted on magnetic beads and concentrated and then an aliquot
of the eluted DNA/RNA is added to PCR reagents which contain the gene-specific primers used to amplify the genetic targets in
the BD MAX PCR Cartridge, if present. The assay also includes a Sample Processing Control. The Sample Processing Control
is present in the Extraction Tube and undergoes the extraction, concentration and amplification steps to monitor for inhibitory
substances, instrument or reagent failure. No operator intervention is necessary once the clinical specimen, the BD MAX Unitized
Reagent Strip and the BD MAX PCR Cartridge are loaded into the BD MAX System. The BD MAX System automates sample
lysis, DNA/RNA extraction and concentration, reagent rehydration, nucleic acid amplification and detection of the target nucleic
acid sequence using real-time polymerase chain reaction (PCR). Amplified targets are detected with hydrolysis probes labeled with
quenched fluorophores. The amplification, detection and interpretation of the signals are done automatically by the BD MAX System.
MAX Enteric Viral Panel-NR - BD
PRINCIPLES OF THE PROCEDURE
Stool specimens are collected from subjects and transported to the laboratory unpreserved in a clean container or preserved in
Cary-Blair transport media. A loop is inserted to the depth of the loop into the specimen and expressed via swirling motion into a
BD MAX Sample Buffer Tube included in the BD MAX Enteric Viral Panel-NR kit. The Sample Buffer Tube is closed with a septum
cap, vortexed and transferred to the BD MAX System. Once the work list is generated and the specimen is loaded on the BD MAX
instrument, along with a BD MAX Enteric Viral Panel Unitized Reagent Strip and PCR Cartridge, the run is started and no further
operator intervention is required. The BD MAX System automates specimen preparation, including target organism lysis, DNA/RNA
extraction and concentration, reagent rehydration, target nucleic acid sequence amplification and detection using real- time PCR.
The interpretation of the signal is performed automatically by the BD MAX System. The assay also includes a Sample Processing
Control that is provided in the Extraction Tube and subjected to extraction, concentration and amplification steps. The Sample
Processing Control monitors for the presence of potential inhibitory substances as well as system or reagent failures. Following
enzymatic viral lysis at elevated temperature, the released nucleic acids are captured by magnetic affinity beads.
The beads, with the bound nucleic acids, are washed and the nucleic acids are eluted. Eluted DNA/RNA is neutralized and
transferred to the Master Mix tubes to rehydrate the PCR reagents. After rehydration, the BD MAX System dispenses a fixed volume
of PCR-ready solution into the BD MAX PCR Cartridge. Microvalves in the BD MAX PCR Cartridge are sealed by the system to
prevent evaporation and amplicon contamination prior to the initiation of reverse transcriptase PCR to convert RNA to cDNA and
subsequent real time PCR.
The amplified DNA targets are detected using hydrolysis (TaqMan®) probes labeled at one end with a fluorescent reporter dye
(fluorophore) and at the other end with a quencher moiety. Probes labeled with different fluorophores are used to detect the
amplicons of the viral targets (Norovirus GI & GII, and Rotavirus A) and the Sample Processing Control amplicons in three
different optical channels of the BD MAX System. When the probes are in their native state, the fluorescence of the fluorophore is
quenched due to its proximity to the quencher. However, in the presence of target DNA, the probes hybridize to their complementary
sequences and are hydrolyzed by the 5′–3′ exonuclease activity of the DNA polymerase as it synthesizes the nascent strand along
the cDNA template. As a result, the fluorophores are separated from the quencher molecules and fluorescence is emitted. The
BD MAX System monitors these signals at each cycle, and interprets the data at the end of the program to report the final results.
REAGENTS AND MATERIALS

      REF                                                  Contents                                             Quantity
                 BD MAX™ Enteric Viral Panel Master Mix (D6)
                                                                                                                 24 tests
                 Dried PCR Master Mix containing DNA polymerase, nucleotides, target and
                                                                                                              (2 x 12 tubes)
                 Sample Processing Control specific probes and primers.
                 BD MAX™ Enteric Viral Panel Unitized Reagent Strips
                 Reagent strips containing all liquid reagents and disposable pipette tips necessary for         24 tests
                 sample processing and DNA/RNA extraction.
                 BD MAX™ Enteric Viral Panel Extraction Tube (D4)
    443987                                                                                                       24 tests
                 Dried extraction reagent containing DNA/RNA magnetic affinity beads, Proteinase K,
                                                                                                              (2 x 12 tubes)
                 and Sample Processing Control
                                                                                                                 24 tests
                 BD MAX™ Enteric Viral Panel Sample Buffer Tube
                                                                                                              (2 x 12 tubes)
                 BD MAX™ Disposable Inoculation Loops
                                                                                                                   30
                 (required for use with the assay)
                 Septum Caps                                                                                       25

                                                                      2
MAX Enteric Viral Panel-NR - BD
EQUIPMENT AND MATERIALS REQUIRED BUT NOT PROVIDED
•   BD MAX™ PCR Cartridges (BD Diagnostic Systems Catalog no. 437519)
•   VWR Multi-Tube Vortexer or equivalent (VWR, Cat. No. 58816-115)
•   Vortex Genie 2 or equivalent (VWR, Cat. No. 58815-234)
•   Nalgene™ Cryogenic Vial Holder (VWR, Cat. No. 66008-783)
•   Rack compatible with a multi-tube vortex mixer (e.g., Cryogenic Vial Holder or equivalent)
•   Lab coat and disposable gloves, powderless
•   Stopwatch or timer
For “Unpreserved” stool specimen type:
•   Dry, clean containers for collection of liquid or soft stool specimens.
For preserved stool specimen type:
•   Cary-Blair transport media (15 mL).
WARNINGS AND PRECAUTIONS
     Danger        H360 May damage fertility or the unborn child.
                   P280 Wear protective gloves/protective clothing/eye protection/face protection.
                   P201 Obtain special instructions before use.
                   P202 Do not handle until all safety precautions have been read and understood.
                   P308+P313 IF exposed or concerned: Get medical advice/attention.
                   P405 Store locked up.
                   P501 Dispose of contents/container in accordance with local/regional/national/international regulations.

•   The BD MAX Enteric Viral Panel-NR is for in vitro diagnostic use only.
•   BD MAX Disposable Inoculating Loops are required for use with the BD MAX Enteric Viral Panel-NR and packaged with the
    assay kit. General loops, regardless of type and volume, are not recommended for use and not validated with the BD MAX
    Enteric Viral Panel-NR performance.
•   Local, state, and federal rules and regulations for notification of reportable diseases are continually updated and include a
    number of organisms that are important for surveillance and outbreak investigations.5,6 Laboratories are responsible for
    following their state and/or local rules pertaining to reportable pathogens and should consult their local and/or state public health
    laboratories for isolate and/or clinical sample submission guidelines.
•   This product can only be used on the BD MAX System.
•   Do not use expired reagents and/or materials.
•   Do not use the kit if the label that seals the outer box is broken upon arrival.
•   Do not use reagents if the protective pouches are open or broken upon arrival.
•   Do not use reagents if desiccant is not present or is broken inside reagent pouches.
•   Do not remove desiccant from reagent pouches.
•   Close protective pouches of reagent promptly with the zip seal after each use. Remove any excess air in the pouches prior
    to sealing.
•   Protect reagents against heat and humidity. Prolonged exposure to humidity may affect product performance.
•   Do not use reagents if the foil has been broken or damaged.
•   Do not mix reagents from different pouches and/or kits and/or lots.
•   Do not interchange or reuse caps, as contamination may occur and compromise test results.
•   Check Unitized Reagent Strips for proper liquid fills (ensure that the liquids are at the bottom of the tubes [refer to Figure 1]).
•   Check Unitized Reagent Strips to ensure that all pipette tips are present (refer to Figure 1).
•   Proceed with caution when using chemical solutions as Master Mix and Extraction tube barcode readability may be altered.
•   Good laboratory technique is essential to the proper performance of this assay. Due to the high analytical sensitivity of this test,
    extreme care should be taken to preserve the purity of all materials and reagents.
•   In case other PCR tests are conducted in the same general area of the laboratory, care must be taken to ensure that the
    BD MAX Enteric Viral Panel-NR, any additional reagents required for testing, and the BD MAX System are not contaminated.
    Avoid microbial, deoxyribonuclease (DNase), and ribonuclease (RNase) contamination of reagents at all times. Gloves must be
    changed before manipulating reagents and cartridge.
•   To avoid contamination of the environment with any target amplicons, do not break apart the BD MAX PCR Cartridge after use.
    The seals in the BD MAX PCR Cartridges are designed to prevent contamination.
•   The laboratory should routinely perform environmental monitoring to minimize the risk of cross-contamination.
•   Performing the BD MAX Enteric Viral Panel-NR outside of the recommended time and temperature ranges recommended
    for specimen transport and storage may produce invalid results. Assays not performed within specified time and temperature
    ranges should be repeated.
•   Additional controls may be tested according to guidelines or requirements of local, state, provincial and/or federal regulations or
    accrediting organizations.

                                                                   3
MAX Enteric Viral Panel-NR - BD
•   Always handle specimens as if they are infectious and in accordance with safe laboratory procedures such as those described
    in CLSI Document M297 and in Biosafety in Microbiological and Biomedical Laboratories.8
•   Wear protective clothing and disposable gloves while handling all reagents.
•   Wash hands thoroughly after performing the test.
•   Do not pipette by mouth.
•   Do not smoke, drink or eat in areas where specimens or kit reagents are being handled.
•   Dispose of unused reagents and waste in accordance with country, federal, provincial, state and local regulations.
•   Consult the BD MAX System User’s Manual9 for additional warnings, precautions and procedures.
STORAGE AND STABILITY
Collected specimens, either unpreserved stool or stool stored in 15 mL Cary-Blair transport media should be kept between 2 °C and
25 °C during transport. Protect against exposure to excessive heat.
Specimen can be stored for up to 120 hours (5 days) at 2–8 °C or for up to 48 hours at 2–25 °C before testing.
BD MAX Enteric Viral Panel Master Mix is stable at 2–25 °C through the stated expiration date. Do not use expired components.
BD MAX Enteric Viral Panel Master Mix Tubes are provided in sealed pouches. To protect product from humidity, immediately
re-seal after opening. Master Mix tubes are stable for up to 14 days at 2–25 °C after initial opening and re-sealing of the pouch.
INSTRUCTIONS FOR USE
Specimen Collection/Transport
In order to obtain an adequate specimen, the procedure for specimen collection must be followed closely. Using a dry, clean
container, liquid or soft stool specimens are collected according to the following procedure:
1. Unpreserved specimens: Transfer the liquid or soft stool specimen to a dry, clean container. Avoid contamination with water or
     urine and avoid mixing toilet paper or soap with the specimen. Label the container and transport to the laboratory according to
     institutional standard operating procedures (refer to the Storage and Stability section).
2. Cary-Blair preserved specimens: Transfer the liquid or soft stool specimen to a 15 mL transport device according to the
     manufacturer’s instructions. Avoid contamination with water or urine and avoid mixing toilet paper or soap with the specimen.
     Label the container and transport to the laboratory according to institutional standard operating procedures (refer to the Storage
     and Stability section).
Specimen Preparation
NOTE: One (1) Sample Buffer Tube, one (1) Septum Cap, one (1) Master Mix Tube (D6), one (1) Extraction Tube (D4) and one (1)
Unitized Reagent Strip are required for each specimen and each External Control to be tested. Set aside the required number of
materials from their protective pouches or boxes. To store opened Master Mix or Extraction Tube pouches, remove excess air and
close using the zip seal.
1. Label a bar-coded BD MAX Sample Buffer Tube (clear cap) with the appropriate specimen identification. Do not obscure, write
    or label over the 2D-barcode.
2. Vortex unpreserved or Cary-Blair preserved specimens at high speed for 15 seconds.
3. Remove the clear cap from the Sample Buffer Tube and inoculate as follows:
    a. Insert a disposable inoculation loop until the entire loop portion is submerged in the specimen. Do not insert beyond the
         loop as any additional stool on the shaft can overload the PCR reaction;
    b. Insert the loaded loop into the Sample Buffer Tube and express the specimen using a swirling motion.
         NOTE: Removal of the entire specimen from the loop is not necessary. The resultant Sample Buffer Tube solution
         should be “tea-stained” in color.
4. Recap the inoculated Sample Buffer Tube using a Septum Cap.
5. Place the Sample Buffer Tube in a rack compatible with a multi-tube vortex mixer, if available (e.g., cryogenic vial holder
    or equivalent).
6. Prepare any additional specimens for testing by repeating Steps 1 through 5, ensuring gloves are clean prior to handling
    additional specimens.
7. Vortex all prepared samples simultaneously at maximum speed for one (1) minute with the multi-tube vortex mixer.
8. Proceed to BD MAX System Operation section to perform testing of the BD MAX Enteric Viral Panel-NR on the
    BD MAX System.
BD MAX System Operation
NOTE: Refer to the BD MAX System User’s Manual9 for detailed instructions (refer to “Operation section”).

                                                                   4
MAX Enteric Viral Panel-NR - BD
NOTE: Testing of the BD MAX Enteric Viral Panel-NR must be performed immediately after the vortexing step above (refer
to “Specimen Preparation”, Step 7). If retesting is necessary, re-vortex sample(s).
1. Power on the BD MAX System (if not already done) and log in by entering  and .
2. Gloves must be changed before manipulating reagents and cartridges.
3. Remove the required number of Unitized Reagent Strips from the BD MAX Enteric Viral Panel-NR kit. Gently tap each Unitized
    Reagent Strip onto a hard surface to ensure that all the liquids are at the bottom of the tubes.
4. Remove from the protective pouches the required number of Extraction Tube(s) and Master Mix Tube(s) from the
    BD MAX Enteric Viral Panel-NR kit.
5. Remove excess air, and close pouches with the zip seal.
6. For each sample to be tested, place one (1) Unitized Reagent Strip on the BD MAX System Rack, starting with Position 1 of
    Rack A.
7. Snap one (1) Extraction Tube (D4) (white foil) into each Unitized Reagent Strip in Position 1 as shown in Figure 1.
8. Snap one (1) BD MAX Enteric Viral Panel Master Mix tube (D6) (green foil) into each Unitized Reagent Strip in Position 2 as
    shown in Figure 1.

                                Figure 1: Snap BD MAX Enteric Viral Panel Extraction tubes and
                            BD MAX Enteric Viral Panel Master Mix tubes into Unitized Reagent Strips

9.    Click on the Run Tab and then the Inventory subtab, enter the kit lot number for the BD MAX Enteric Viral Panel-NR and master
      mix lot number for the BD MAX Enteric Viral Panel-NR (for lot traceability) by either scanning the barcode with the scanner or by
      manual entry.
      NOTE: Repeat step 9 each time a new kit lot is used.
10.   Navigate to the Worklist. Using the pull down menu select .
11.   Select the appropriate kit lot number (found on the outer box of the BD MAX Enteric Viral Panel-NR) and master mix lot number
      for BD MAX Enteric Viral Panel-NR from the pull down menu.
12.   Enter the BD MAX Enteric Viral Panel Sample Buffer Tube ID, Patient ID and Accession Number (if applicable) into the Worklist,
      either by scanning the barcode with the scanner or by manual entry.
13.   Repeat step 12 for all remaining Sample Buffer Tubes.
14.   Place the Sample Buffer Tubes in the BD MAX System Rack(s) corresponding to the Unitized Reagent Strips assembled in
      steps 6 to 8.
      NOTE: Place the Sample Buffer Tubes into the specimen rack with 1D barcode labels facing outward (this makes
      scanning Sample Buffer Tubes easier during sample login).
15.   Place the required number of BD MAX PCR Cartridge(s) into the BD MAX System (refer to Figure 2):
      •    Each BD MAX PCR Cartridge accommodates 1 run of up to 12 samples for a total of 12 samples.
      •    The BD MAX System will automatically select the position and row on the BD MAX PCR Cartridge for each run.
      •    BD MAX PCR Cartridges are used on a per-run AND rack basis (1 run per cartridge and 1 cartridge per rack).
      •    To maximize use of BD MAX PCR Cartridges, using 2000 Sample Mode, select Run Wizard under the Worklist tab for
           lane assignments.
      •    Consult the BD MAX System User’s Manual9 for more details.

                                                                   5
Figure 2: Load BD MAX PCR Cartridges

16. Load rack(s) into the BD MAX System (refer to Figure 3).

                                                   Side A                  Side B
                                         Figure 3: Load Rack(s) into the BD MAX System

17. Close the BD MAX System lid and click the  to begin processing.
18. At the end of the run, check results immediately or store Sample Buffer Tubes at 2–8 °C for up to 5 days (120 hours)
    OR at 2–25 °C for a maximum of 48 hours until the results are checked.
NOTE: If a septum cap was damaged during the run, replace it with a new one before storing the sample.
NOTE: Prepared BD MAX Sample Buffer Tubes can be stored at 2–8 °C for a maximum of 120 hours (5 days) OR at 2–25 °C
for a maximum of 48 hours. When an Indeterminate (IND), Unresolved (UNR), or Incomplete (INC) result is obtained, or
when an External Control failure occurs, a repeat test from the prepared Sample Buffer Tube must be performed within this
timeframe (refer to Repeat Test Procedure section).
QUALITY CONTROL
Quality control procedures monitor the performance of the assay. Laboratories must establish the number, type and frequency
of testing control materials according to guidelines or requirements of local, provincial, state, federal and/or country regulations
or accreditation organizations in order to monitor the effectiveness of the entire analytical process. For general Quality Control
guidance, the user may wish to refer to Clinical Laboratory Standards Institute documents MM310 and EP12.11
1. External Control materials are not provided by BD. External Positive and Negative Controls are not used by the BD MAX
     System software for the purpose of sample test result interpretation. External Controls are treated as if they were patient
     samples. (Refer to Table 2 for the interpretation of External Control assay results.)
2. One (1) External Positive Control and one (1) External Negative Control should be run at least daily until adequate process
     validation is achieved on the BD MAX System in each laboratory setting. Reduced frequency of control testing should be in
     accordance with applicable regulations.
3. The External Positive Control is intended to monitor for substantial reagent failure. The External Negative Control is intended to
     detect reagent or environmental contamination (or carry-over) by target nucleic acids.

                                                                  6
4.     Various types of External Controls are recommended to allow the user to select the most appropriate for their laboratory quality
       control program.
       a. External Negative Control: Commercially available control material or a previously characterized sample known to be
           negative. BD recommends that the External Negative Control be prepared prior to the External Positive Control in order to
           reduce the potential for contamination as a result of control preparation.
       b. External Positive Control: Commercially available control materials, such as the ZeptoMetrix® strains listed below (Refer to
           Table 1), or previously characterized samples known to be positive.

                                   Table 1: Commercially Available Strains for External Positive Control
                                  External Positive Control Strain                             Part Number
                                              Rotavirus A                         ZeptoMetrix 0810041CF or 0810281CF
                                  Recombinant Norovirus GI or GII                 ZeptoMetrix 0810086CF or 0810087CF

       For the preparation of External Control suspension, it is recommended that each viral culture fluid be diluted with a 1:10 dilution
       in TE buffer. Inoculate 5 μL of the viral suspension into the corresponding Sample Buffer Tube. Process and test as a sample
       (refer to the Specimen Preparation and BD MAX System operation sections).
5.     All External Controls should yield the expected results (positive for External Positive Control, negative for External Negative
       Control) and no failed external controls (Unresolved, Indeterminate, Incomplete results).
6.     An External Negative Control that yields a positive test result is indicative of a specimen handling and/or contamination event.
       Review the specimen handling technique to avoid mix-up and/or contamination. An External Positive Control that yields a
       negative result is indicative of a specimen handling/preparation problem. Review the specimen handling/preparation technique.
7.     An External Control that yields an Unresolved, Indeterminate or Incomplete test result is indicative of a reagent or a BD MAX
       System failure. Check the BD MAX System monitor for any error messages. Refer to the “System Error Summary” section of
       the BD MAX System User’s Manual9 for interpretation of warning and error codes. If the problem persists, use reagents from an
       unopened pouch or use new BD MAX Enteric Viral Panel-NR kit.
8.     Each BD MAX Enteric Viral Panel Extraction Tube contains a Sample Processing Control which is an Armored RNA construct
       containing a synthetic target RNA sequence. The Sample Processing Control is extracted, eluted and amplified along with any
       DNA/RNA present in the processed specimen, ensuring predictivity of the assay. The Sample Processing Control monitors
       the efficiency of DNA/RNA capture, washing and elution during the sample processing steps, as well as the efficiency of RNA
       conversion to DNA and DNA target amplification and detection during PCR analysis. If the Sample Processing Control result
       fails to meet the acceptance criteria, the result of the sample will be reported as Unresolved; however, any positive (POS)
       assay results will be reported and no targets will be called negative (NEG). This will be reported on a per Master Mix basis. An
       Unresolved result is indicative of specimen-associated inhibition or reagent failure. Repeat any sample reported as Unresolved
       according to the “Repeat Test Procedure” section below.
RESULTS INTERPRETATION
Results are available on the  tab in the  window on the BD MAX System monitor. The BD MAX System
software automatically interprets test results. Results are reported for each of the analytes and for the Sample Processing Control. A
test result may be called as NEG (negative), POS (positive) or UNR (unresolved) based on the amplification status of the target and
of the Sample Processing Control. IND (Indeterminate) or INC (Incomplete) results are due to BD MAX System failure. In the case
of a partial UNR, where one target has a POS result and the other target has a UNR result, the target with a UNR result will not be
called NEG. This will be reported on a per Master Mix basis.
BD MAX Enteric Viral Panel-NR results interpretation is described below in Table 2.

                                        Table 2: BD MAX Enteric Viral Panel-NR Result Interpretation
    ASSAY RESULT REPORTED                                                    INTERPRETATION OF RESULT
              NoV POS                                                       Norovirus GI or GII RNA detected
              RoV POS                                                             Rotavirus A RNA detected
              NoV NEG                                    No Norovirus RNA detected and Sample Processing Control detected
              RoV NEG                                   No Rotavirus A RNA detected and Sample Processing Control detected
                                                  No Norovirus GI or GII RNA detected and no Sample Processing Control is detected
              NoV UNR
                                                                (indicative of an inhibitory specimen or reagent failure)
                                                               No Rotavirus A RNA detected and no Sample Processing
              RoV UNR
                                                        Control detected (indicative of an inhibitory specimen or reagent failure)
                                                                      Indeterminate due to BD MAX System failure
         Indeterminate (IND)
                                                                             (with Warning or Error Codesa)
                                                                                       Incomplete Run
          Incomplete (INC)
                                                                                (with Warning or Error Codesa)
a   Refer to “Troubleshooting” section of the BD MAX System User’s Manual9 for interpretation of warning and error codes.

                                                                            7
REPEAT TEST PROCEDURE
NOTE: Sufficient volume is available for one repeat test from the Sample Buffer Tube. For Sample Buffer Tubes stored at
2–25 °C, retesting must be performed within 48 h following the initial Sample Buffer Tube inoculation with the specimen.
Alternatively, for Sample Buffer Tubes stored at 2–8 °C, retesting may be performed within 120 hours (5 days) following
the initial Sample Buffer Tube inoculation with the specimen. The remaining stool specimen may also be used for repeat
testing within 120 hours (5 days) of collection if stored at 2–8 °C or within 48 hours if stored at 2–25 °C.
NOTE: New samples may be tested in the same run with repeat samples.
UNRESOLVED RESULT
Unresolved results may be obtained in the event that sample-associated inhibition or a reagent failure prevents proper target or
Sample Processing Control amplification. If the Sample Processing Control does not amplify, the sample will be reported as UNR;
however, any positive (POS) assay results will be reported and all other targets will be called UNR.
The BD MAX System reports results for each target individually and a UNR result may be obtained for one or more BD MAX Enteric
Viral Panel-NR targets. In the case of a complete UNR, where all targets have a UNR result, it is necessary to repeat the test. In
the case of a partial BD MAX Enteric Viral Panel-NR UNR result, when one target has a POS result and the other target has a UNR
result, the test should be repeated as described above. In rare cases, discrepant results may be observed when a repeat test is run
for those targets that were initially reported as POS. In that case, any positive result is retained.
Sample(s) can be repeated from their corresponding Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s)
for one (1) min and restart from the “BD MAX System Operation” section. The remaining stool specimen may also be used for repeat
testing with a new Sample Buffer Tube within the timeframes defined above. Restart from the Specimen Preparation section.
INDETERMINATE RESULT
Indeterminate results may be obtained in the event that a System failure occurs. Sample(s) can be repeated from their corresponding
Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s) for one (1) minute and restart following the
“BD MAX System Operation” section. The remaining stool specimen, with a new Sample Buffer Tube, may also be used for repeat
testing within the timeframe defined above. Restart from the Specimen Preparation section. For the interpretation of warning or error
code messages, refer to the BD MAX User’s Manual9 (Refer to “Troubleshooting” section).
INCOMPLETE RESULT
Incomplete results may be obtained in the event that the Specimen Preparation or the PCR failed to complete. Sample(s) can be
repeated from their corresponding Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s) for one (1) minute
and restart following “BD MAX System Operation” section. The remaining stool specimen, with a new Sample Buffer Tube, may also
be used for repeat testing within the timeframe defined above. Restart from the Specimen Preparation section. For the interpretation of
warning or error code messages, refer to the BD MAX System User’s Manual9 (Refer to “Troubleshooting” section).
EXTERNAL CONTROL FAILURE
External Controls should yield expected results when tested. If samples have to be repeated due to an incorrect External Control
result, they should be repeated from their Sample Buffer Tubes along with freshly prepared External Controls within the timeframes
defined above. Vortex the samples for one (1) minute and restart following the “BD MAX System Operation” section.
CULTURING OF SPECIMENS
Culture and identification of organisms from positive specimens should be performed per laboratory procedures.
LIMITATIONS OF THE PROCEDURE
•   This product should only be used with the BD MAX System.
•   This product is intended for use only with unpreserved and Cary-Blair preserved human stool specimens. Stool specimens from
    rectal swabs or fixed stools have not been validated with the BD MAX Enteric Viral Panel-NR.
•   Erroneous results may occur from improper specimen collection, handling, storage, technical error, specimen mix-up or because
    the number of organisms in the specimen is below the analytical sensitivity of the test.
•   If the BD MAX Enteric Viral Panel-NR result is IND, INC, or UNR (for one or more targets) then the test should be repeated.
•   A BD MAX Enteric Viral Panel-NR positive result does not necessarily indicate the presence of viable organisms. It does
    however indicate the presence of target DNA/RNA.
•   Mutations or polymorphisms in primer- or probe-binding regions may affect detection of new or unknown target variants,
    resulting in a false negative result with the BD MAX Enteric Viral Panel-NR.
•   The BD MAX Enteric Viral Panel-NR simultaneously detects Norovirus GI & GII, and does not differentiate between the species.
•   As with all PCR-based tests, extremely low levels of target below the LoD of the assay may be detected, but results may not
    be reproducible.
•   False negative results may occur due to loss of nucleic acid from inadequate collection, transport or storage of specimens, or
    due to inadequate viral lysis. The Sample Processing Control has been added to the test to aid in the identification of samples
    that contain inhibitors to PCR amplification and as a control for reagent integrity and of the assay system as a whole. The
    Sample Processing Control does not indicate if nucleic acid has been lost due to inadequate collection, transport or storage of
    samples, or whether viral capsids have been adequately lysed.
•   Results from the BD MAX Enteric Viral Panel-NR should be used as an adjunct to clinical observations and other information
    available to the physician.

                                                                   8
•   As with all in vitro diagnostic tests, positive and negative predictive values are highly dependent on prevalence. The BD MAX
    Enteric Viral Panel-NR performance may vary depending on the prevalence and population tested.
•   BD MAX Enteric Viral Panel-NR results may or may not be affected by concurrent medical therapy, which may reduce the
    amount of target present.
•   The Sample Buffer Tube has not been designed to support organism viability. If culture is necessary it must be performed from
    the original specimen.
•   The performance of this test has not been established for monitoring treatment of Norovirus GI & GII, and Rotavirus A infections.
•   This test is a qualitative test and does not provide quantitative values nor indicate the quantity of organisms present.
•   The performance of this test has not been evaluated for immunocompromised individuals or for patients without symptoms of
    gastrointestinal infection.
•   The effect of interfering substances has only been evaluated for those listed in this labeling. Potential interference has not been
    evaluated for substances other than those described in the Interference section below. Interference by substances other than
    those described in the interference section below could lead to erroneous results.
•   BD MAX Enteric Viral Panel-NR assay performance has not been evaluated in individuals who have received the Rotavirus A
    vaccine, which is known to react with this assay.
•   Cross-reactivity with organisms other than those listed in the Analytical Specificity section below has not been evaluated.
EXPECTED VALUES
In the BD MAX Enteric Viral Panel-NR trial, reportable results were obtained from six (6) geographically diverse clinical centers. The
study was performed from two (2) stool specimen types, Cary-Blair preserved and unpreserved. The number and percentage of
positive cases per target, as determined in the BD MAX Enteric Viral Panel-NR trial, are presented in Table 3.

Table 3: Expected Values Rate based on BD MAX Enteric Viral Panel-NR Assay Results by Specimen Type, Site and Overall
        Specimen Type                           Site                           Norovirus                         Rotavirus
                                                ALB                            7.7% (4/52)                      0.0% (0/52)
                                                CIN                         15.0% (59/394)                    6.9% (27/394)
      Cary-Blair Preserved                      JHU                          9.8% (13/132)                     4.5% (6/132)
                                                POR                          3.9% (14/359)                     1.9% (7/359)
                                                Total                        9.6% (90/937)                    4.3% (40/937)
                                                CAL                          5.4% (31/574)                     1.0% (6/574)
                                                CIN                             0.0% (0/3)                      33.3% (1/3)
                                                JHU                           13.8% (9/65)                      0.0% (0/65)
          Unpreserved
                                                LUR                          10.1% (10/99)                      5.1% (5/99)
                                                POR                           23.1% (3/13)                      7.7% (1/13)
                                                Total                        7.0% (53/754)                    1.7% (13/754)
                                                ALB                            7.7% (4/52)                      0.0% (0/52)
                                                CAL                          5.4% (31/574)                     1.0% (6/574)
                                                CIN                         14.9% (59/397)                    7.1% (28/397)
           Combined                             JHU                         11.2% (22/197)                     3.0% (6/197)
                                                LUR                          10.1% (10/99)                      5.1% (5/99)
                                                POR                          4.6% (17/372)                     2.2% (8/372)
                                                Total                      8.5% (143/1,691)                  3.1% (53/1,691)

                                                                  9
PERFORMANCE CHARACTERISTICS
Clinical Performance characteristics of the BD MAX Enteric Viral Panel-NR were determined in a multi-site investigational study.
The study involved a total of six (6) geographically diverse clinical centers where stool specimens were collected as part of routine
patient care, enrolled into the trial, and tested with the BD MAX Enteric Viral Panel-NR. Specimens were obtained from pediatric or
adult patients suspected of acute gastroenteritis, enteritis or colitis, for whom diagnostic procedures were indicated and/or ordered
by a healthcare provider. The reference method for the prospective specimens was a combination of two (2) sets of alternate PCRs
and bi-directional sequencing for one (1) PCR. All prospective specimens were tested fresh (stored 2–8 °C and within 5 days of
collection) with the BD MAX Enteric Viral Panel-NR Assay, but frozen prior to testing with the reference method. For retrospective
specimens, the historical results were recorded at the collection site. All retrospective specimens were frozen prior to testing on the
BD MAX Enteric Viral Panel-NR Assay and the reference method. The historical results were confirmed using an alternate PCR
assay and bi-directional sequencing in order to confirm the presence of the target DNA.
A total of 1,873 prospective specimens (1,055 Cary-Blair preserved and 818 unpreserved) and 366 retrospective specimens
(136 Cary-Blair preserved and 230 unpreserved) were enrolled in the clinical evaluation for a total of 2,239 specimens enrolled.
Table 4 describes the number of compliant specimens enrolled by patient age and specimen type with a total of 2,148 compliant
specimens overall. Tables 5 and 6 describe the performance characteristics of the BD MAX Enteric Viral Panel-NR that were
observed during the clinical trial.

                        Table 4: Compliant Clinical Trial Enrollment Summary by Age Group and Specimen Type

             Age Group                        Cary-Blair Preserved                       Unpreserved                            Combined
             0–1 month                                  4                                      0                                     4
          1 month to 2 years                          188                                    112                                   300
                2–12                                  228                                    153                                   381
               13–18                                  117                                     66                                   183
               19–21                                   20                                     21                                    41
              Over 21                                 568                                    640                                  1,208
              Unknown                                  21                                     10                                    31
                Total                                1,146                                  1,002                                 2,148

Norovirus Performance Results
For the Cary-Blair preserved specimen type, the BD MAX Enteric Viral Panel-NR identified 92.5% and 99.2% of the Norovirus
prospective positive and negative specimens, respectively, and 100% and 99.1% of the retrospective positive and negative
specimens, respectively. For the unpreserved specimen type, the BD MAX Enteric Viral Panel-NR identified 90.7% and 99.6% of the
Norovirus prospective positive and negative specimens, respectively and 94.6% and 100% of the Norovirus retrospective positive
and negative specimens, respectively. Refer to Table 5.

                                  Table 5. Norovirus- Performance Results per Specimen Type and Origin
                                                                                                                RM
                 Specimen Type                          Specimen Origin              BD MAX                                                Total
                                                                                                         P               N
                                                                                   P                     74              7a                81
                                                           Prospective
               Cary-Blair Preserved                                                N                     6b             835                841
                                                             (Fresh)
                                                                                  Total                  80             842                922
                                                         PPA (95% CI): 92.5% (84.6%, 96.5%)
                                                         NPA (95% CI): 99.2% (98.3%, 99.6%)
                                                                                   P                     6               1                  7
                                                         Retrospective
               Cary-Blair Preserved                                                N                     0              105                105
                                                           (Frozen)
                                                                                  Total                  6              106                112
                                                          PPA (95% CI): 100% (61%, 100%)
                                                         NPA (95% CI): 99.1% (94.8%, 99.8%)
                                                                                   P                     39              3                  42
                                                          Prospective
                   Unpreserved                                                     N                      4             694                698
                                                            (Fresh)
                                                                                  Total                  43             697                740
                                                         PPA (95% CI): 90.7% (78.4%, 96.3%)
                                                         NPA (95% CI): 99.6% (98.7%, 99.9%)
                                                                                   P                     35              0                  35
                                                         Retrospective
                   Unpreserved                                                     N                     2              58                  60
                                                           (Frozen)
                                                                                  Total                  37             58                  95
                                                         PPA (95% CI): 94.6% (82.3%, 98.5%)
                                                         NPA (95% CI): 100% (93.8%, 100%)
a
    7/7 Specimens were available to be tested in discrepant analysis and 4/7 tested positive for Norovirus with the FilmArray™ Gastrointestinal Panel.
b
    6/6 Specimens tested negative for Norovirus during discrepant analysis with the FilmArray Gastrointestinal Panel.

                                                                            10
Rotavirus Performance Results
For the Cary-Blair preserved specimen type, the BD MAX Enteric Viral Panel-NR identified 100% and 99.2% of the Rotavirus
prospective positive and negative specimens, respectively, and 100% and 98.7% of the Rotavirus retrospective positive and
negative specimens, respectively. For the unpreserved specimen type, the BD MAX Enteric Viral Panel-NR identified 100%
and 99.9% of the Rotavirus prospective positive and negative specimens, respectively and 100% and 97.9% of the Rotavirus
retrospective positive and negative specimens, respectively. Refer to Table 6.

                                   Table 6. Rotavirus- Performance Results per Specimen Type and Origin
                                                                                                       RM
      Specimen Type           Specimen Origin                BD MAX                                                                        Total
                                                                                          P                         N
                                                                P                    31                            7a                        38
                                   Prospective
    Cary-Blair Preserved                                        N                     0                           888                       888
                                     (Fresh)
                                                             Total                   31                           895                       926
                                                           PPA (95% CI): 100% (89%, 100%)
                                                          NPA (95% CI): 99.2% (98.4%, 99.6%)
                                                                P                    38                             1                        39
                                 Retrospective
    Cary-Blair Preserved                                        N                     0                            76                        76
                                   (Frozen)
                                                             Total                   38                            77                       115
                                                          PPA (95% CI): 100% (90.8%, 100%)
                                                           NPA (95% CI): 98.7% (93%, 99.8%)
                                                                P                    11                            1                        12
                                   Prospective
        Unpreserved                                             N                     0                           735                       735
                                     (Fresh)
                                                             Total                   11                           736                       747
                                                          PPA (95% CI): 100% (74.1%, 100%)
                                                          NPA (95% CI): 99.9% (99.2%, 100%)
                                 Retrospective                  P                    56                             1                        57
        Unpreserved                (Frozen)                     N                     0                            47                        47
                                                             Total                   56                            48                       104
                                                          PPA (95% CI): 100% (93.6%, 100%)
                                                          NPA (95% CI): 97.9% (89.1%, 99.6%)
a
    7/7 Specimens were available to be tested in discrepant analysis and 4/7 tested positive for Rotavirus with the FilmArray Gastrointestinal Panel.

NON-REPORTABLE RATE
The initial unresolved rate of the 1,873 prospective specimens evaluated in this study was 0.8% of the Cary-Blair preserved and
1.8% of the unpreserved specimens. The unresolved rate following a valid repeat test was only 0.1% of the Cary-Blair preserved
specimens and 1.0% of the unpreserved specimens (refer to Table 7).
Of the 1,873 prospective specimens evaluated in this study, 0.5% of the Cary-Blair preserved and 2.2% of the unpreserved initially
reported as Indeterminate. Following a valid repeat test, 0.1% of the Cary-Blair preserved and none of the unpreserved specimens
remained Indeterminate (refer to Table 7).
Of the 1,873 prospective specimens evaluated in this study, 0.1% of the Cary-Blair preserved and 0.3% of the unpreserved
specimens initially reported as Incomplete. Following a valid repeat test, 0.1% of the Cary-Blair preserved and none of the
unpreserved specimens remained Incomplete (refer to Table 7).

                            Table 7: Non-reportable Rates for Combined Target by Specimen Type and Overall
     Combined
                           Unresolved Rate                 Indeterminate Rate                 Incomplete Rate                       Total Rate
      Target
     Specimen        Initial EVP       Final EVP       Initial EVP       Final EVP       Initial EVP        Final EVP      Initial EVP       Final EVP
       Type           (95% CI)          (95% CI)        (95% CI)          (95% CI)        (95% CI)           (95% CI)       (95% CI)          (95% CI)
                        0.8%              0.1%            0.5%             0.1%             0.1%             0.1%             1.4%             0.3%
     Cary-Blair
                       9/1,085           1/1,076         5/1,085          1/1,076          1/1,085          1/1,076         15/1,085          3/1,076
     Preserved
                    (0.4%, 1.6%)      (0.0%, 0.5%)    (0.2%, 1.1%)     (0.0%, 0.5%)     (0.0%, 0.5%)     (0.0%, 0.5%)     (0.8%, 2.3%)     (0.1%, 0.8%)
                        1.8%              1.0%            2.2%             0.0%             0.3%             0.0%             4.3%             1.0%
    Unpreserved        18/997            10/982          22/997            0/982            3/997            0/982           43/997           10/982
                    (1.1%, 2.8%)      (0.6%, 1.9%)    (1.5%, 3.3%)     (0.0%, 0.4%)     (0.1%, 0.9%)     (0.0%, 0.4%)     (3.2%, 5.8%)     (0.6%, 1.9%)
                        1.3%              0.5%            1.3%             0.0%             0.2%             0.0%             2.8%             0.6%
     Combined         27/2,082          11/2,058        27/2,082          1/2,058          4/2,082          1/2,058         58/2,082         13/2,058
                    (0.9%, 1.9%)      (0.3%, 1.0%)    (0.9%, 1.9%)     (0.0%, 0.3%)     (0.1%, 0.5%)     (0.0%, 0.3%)     (2.2%, 3.6%)     (0.4%, 1.1%)

                                                                             11
Analytical Inclusivity
A variety of BD MAX Enteric Viral Panel-NR assay target strains were included in this study. Strain selection criteria included
prevalence, serotype and geographic location, where appropriate. Twenty-seven (27) strains were tested, including strains from
public collections and well-characterized clinical isolates.
Inclusivity testing included 22 strains of Norovirus GI & GII and five (5) strains of Rotavirus. The strains were tested at ≥ 3 x LoD
(Limit of Detection) of the corresponding strain in unpreserved stool matrix. The BD MAX Enteric Viral Panel-NR correctly identified
all strains tested upon initial testing. Rotavirus strain WISC2 VR-2517 required testing above 3x LoD, and was resolved at 20x
LoD. In silico analysis predicts that most strains of all genotypes will be detected, though some variant strains may be detected
with reduced sensitivity or may not be detected due to inefficient amplification. For Norovirus in silico analysis, three (3) sequences
showed more than one mismatch, two GI.3 variants and one GI.7 variant. Some Norovirus sequences showed more than two
mismatches, one GII.3 variant, one GII.4 variant, one GII.6 variant and one GII.12 variant. These mutations could affect the
detection of these variants. For Rotavirus A in silico analysis, there were five (5) variants that had more than three (3) mismatches.
There were ten (10) Rotavirus A sequences that were truncated by four (4) nucleotides. These mutations could affect the detection
of these variants.
Noroviruses are genetically diverse. In silico analysis predicts that most strains, including NoV GI.3, GII.P16_GII.4, GII.P16_GII.2,
and GII.Pe_GII.2, may be detected (refer to Table 8). Some variant strains may be detected with reduced sensitivity, or may not be
detected due to inefficient amplification.

                                                   Table 8: Analytical Inclusivity Strains Tested
                         Virus                                             Type                              Strains Tested
                                                                             3                                     0a
                     Norovirus GI                                           4                                      1
                                                                            6                                      1
                                                                            1                                      2
                                                                            2                                      1
                                                                            3                                      2
                                                                            4                                      8
                                                                            6                                      3
                     Norovirus GII                                          12                                     2
                                                                            17                                     1
                                                                         P16-GII.2                                 0a
                                                                         P16-GII.4                                 0a
                                                                         Pe_GII.2                                  0a
                                                                         Unknown                                   1
                       Rotavirus                                            A                                      5
a   Genotypes with unavailable strains were evaluated with in silico binding analysis.

Analytical Sensitivity
The analytical sensitivity (Limit of Detection or LoD) for the BD MAX Enteric Viral Panel-NR was determined as follows: Each target
organism was prepared and quantified prior to inclusion in this study. Individual inoculating loops were dipped into each organism
preparation and were then transferred to a Sample Buffer Tube already containing fecal matrix (preserved or unpreserved) that
was pre-determined to be negative for all the targets detected by the BD MAX Enteric Viral Panel-NR. Each organism was tested
with a minimum of 20 replicates per sample type (preserved or unpreserved), by 2 operators, using 3 different production lots of
the BD MAX Enteric Viral Panel. The LoD for a specific organism was confirmed by testing at least 20 additional replicates at the
determined LoD concentration. Analytical sensitivity (LoD), defined as the lowest concentration at which greater than or equal to
95% of all replicates are expected to test positive (refer to Table 9).

                             Table 9: BD MAX Enteric Viral Panel-NR Limit of Detection for Individual Targets
                                                                                         Unpreserved LoD      Cary-Blair Preserved LoD
           Target Organism                              Strain
                                                                                          (cp/mL in stool)        (cp/mL in stool)
                                                          GI                                 6.28E+06                 4.71E+06
                Norovirus
                                                          GI                                 2.49E+05                 2.49E+05
                                                          WA                                 6.46E+03                 1.29E+04
                Rotavirus
                                                         Va70                                1.16E+04                 5.82E+03

Analytical Specificity (Cross-Reactivity and Exclusivity)
The BD MAX Enteric Viral Panel-NR was performed on samples containing phylogenetically related species and other organisms
(bacteria, viruses, parasites and yeast) likely to be found in stool specimens. The bacterial cells, yeasts, parasites and viruses were
tested in the Sample Buffer Tube at ≥ 106 CFU, cells or genome equivalents/mL in stool, or ≥ 105 PFU/mL in stool or TCID50/mL in
stool. Overall, 112 organisms were tested and no cross-reactivity was observed.

                                                                             12
Interfering Substances
Thirty-two (32) biological and chemical substances that may occasionally be present in stool specimens were evaluated for potential
interference with the BD MAX Enteric Viral Panel-NR. Included in this study was an Antibiotics Mixture, which consisted of a
combination of 7 different antibiotics or analgesics tested simultaneously. These antibiotics or analgesics included Naproxen sodium,
Ceftriaxone disodium, Erythromycin, Metronidazole, Sulfamethoxazole, Tetracycline hydrochloride, and Trimethoprim. RotaTeq
vaccine also yielded positive results as expected because the vaccine can be present in the stool up to 9 days post vaccination.12
Results demonstrated no reportable interference with any other substance tested (refer to Table 10).

       Table 10: Endogenous and Commercial Exogenous Substances Tested with the BD MAX Enteric Viral Panel-NR
      Brand Name or Description                  Result                           Brand Name or Description                   Result
               Fecal Fat                              NI                               Spermicial Lubricant                     NI
                 Mucus                                NI                              Suppository (Glycerin)                    NI
           Whole Human Blood                          NI                                     Vagisil                            NI
          Hydrocortisone Cream                        NI                                    Laxatives                           NI
           Antiseptic Towelettes                      NI                              Anti-Diarrheal (liquid)                   NI
            Enema; Mineral Oil                        NI                               Anti-Diarrheal (pill)                    NI
             Hemorrhoidal Gel                         NI                                Antibiotics Mixture                     NI
              Nystatin Cream                          NI                                     Antacids                           NI
             Topical Antibiotic                       NI                      Non-Steroidal Anti-Inflamatory (NSAID)            NI
NI: No reportable interference with the BD MAX Enteric Viral Panel-NR.

In addition, microorganisms that may be endogenously present in stool specimens were evaluated for potential interference with
the BD MAX Enteric Viral Panel-NR. Five (5) organisms were tested at high concentration (1 x106 cells/mL of stool). Results
demonstrated no reportable interference with any microorganism tested (refer to Table 11).

                    Table 11: Microorganisms Tested for Interference with the BD MAX Enteric Viral Panel-NR
                                   Microorganism                                                                Result
                             Salmonella typhimurium                                                               NI
                                   Escherichia coli                                                               NI
                                   Proteus vulgaris                                                               NI
                              Enterococcus faecalis                                                               NI
                         Peptostreptococcus anaerobius                                                            NI
NI: No reportable interference with the BD MAX Enteric Viral Panel-NR.
Mixed Infection / Competitive Interference
The mixed infection/competitive interference study was designed to evaluate the ability of the BD MAX Enteric Viral Panel-NR to
detect low positive results in the presence of other targets at high concentrations. Organisms (Norovirus, and Rotavirus) for Master
Mix D6 were prepared at the 95th percentile observed in the clinical trial to simulate a high clinical load to serve as high targets in
the BD MAX Enteric Viral Panel Sample Buffer Tube. The BD MAX Enteric Viral Panel-NR analyte absent from the high targets mix
was spiked into the Sample Buffer Tube at a concentration 2x their respective LoD representing a low load target along with 5 μL
of unpreserved stool and tested to simulate mixed infections. In the presence of high loads, all organisms corresponding to their
respective simulated mixed infection preparations were successfully detected by the BD MAX Enteric Viral Panel-NR.

                                                                         13
Freeze/Thaw Study
This study was designed to evaluate multiple freeze/thaw cycles at varied LoD levels (1.99x, 4x and 10x). Based on the study
results, three (3) freeze/thaw cycles do not affect the performance of BD MAX Enteric Viral Panel-NR. The BD MAX Enteric Viral
Panel-NR was able to detect 100% proportion positive for all enteric viral targets spiked in preserved and unpreserved, negative,
clinical stool specimens before and after undergoing multiple freeze/thaw cycles.

                                                Table 12: Summary of Freeze/Thaw Study Results
                                                     Target Positive Matrix Proportion Positive (%) and total number of samples tested
                    Condition
                                                                                   (across all LoD levels)
       Matrix         Freeze/Thaw Cycles                         Norovirus GII                                       Rotavirus Va70
                                                                      100                                                  100
                         0 (fresh/baseline)
                                                                     60/60a                                               60/60a
                                                                      100                                                  100
                                 1
                                                                     60/60                                                60/60
 Unpreserved
                                                                      100                                                  100
                                 2
                                                                     60/60                                                60/60
                                                                      100                                                  100
                                 3
                                                                     60/60                                                60/60
                                                                      100                                                  100
                         0 (fresh/baseline)
                                                                     60/60                                                60/60
                                                                      100                                                  100
                                 1
                                                                     60/60a                                               60/60a
   Preserved
                                                                      100                                                  100
                                 2
                                                                     60/60a                                               60/60a
                                                                      100                                                  100
                                 3
                                                                     60/60a                                               60/60a
a Proportion    positive rates were calculated based on total sample number after removal of IND/UNR results, which were excluded from the analysis.

Precision
Within–laboratory precision was evaluated for the BD MAX Enteric Viral Panel-NR at one site. Testing was performed over 12 days,
with two runs per day (one each by 2 operators), for a total of 24 runs. Five specific target organisms, at different concentrations,
were used to create the panel members for this study. The panel members contained Norovirus and Rotavirus.
The following values were used as spike levels for the target organisms contained in each panel member:
•   Moderate Positive (MP): 2.99x LoD
•   Low Positive (LP): 1.99x LoD
•   True Negative (TN): No target
Each panel member was spiked with negative unpreserved stool matrix. True negative samples contained no target. Results are
summarized by target and concentration in Table 13.

                          Table 13: Precision Study Result Using One Lot of the BD MAX Enteric Viral Panel-NR
                                                                                   Agreement with Expected Results
                       Category                                         Norovirus                                  Rotavirus
                                                                         (95% CI)                                  (95% CI)
                                                                            100                                       99.5
                          TNa                                             192/192                                   191/192
                                                                        (98.0, 100)                               (97.1, 99.9)
                                                                            100                                       100
                           LP                                              48/48                                     48/48
                                                                        (92.6, 100)                               (92.6, 100)
                                                                            100                                       100
                          MP                                               48/48                                     48/48
                                                                        (92.6, 100)                               (92.6, 100)
aFor
       the True Negative (TN) category, the reported agreement indicates the percent of negative results.

                                                                            14
Reproducibility
The Site-to-Site reproducibility study was performed at three (3) clinical sites using one (1) reagent lot. Two (2) operators performed
2 runs per day, over five (5) distinct days (consecutive or not), for a total of 30 runs. The panels used were the same as described
under the Precision heading, above.
The site-to-site reproducibility percent agreements were 100% for the TN for all targets, and ranged from 97.8 to 100% and
96.7 to 100% for the LP and MP respectively. Results are summarized in Table 14. The quantitative reproducibility results across
sites and by target are presented in Table 14. Ct.Score and the Cycle EP, internal criteria used to determine a final assay result, was
selected as a means of assessing quantitative assay reproducibility. Mean Ct.Score and the mean Cycle EP values with variance
components (SD and % CV) are shown in Table 15.

                  Table 14. Site-to-site Reproducibility Results Using One Lot of the BD MAX Enteric Viral Panel-NR
                                                                                     Agreement with Expected Results
                      Category                                          Norovirus                                        Rotavirus
                                                                         (95% CI)                                        (95% CI)
                                                                            100                                              100
                         TNa                                              360/360                                          360/360
                                                                        (98.9, 100)                                      (98.9, 100)
                                                                            100                                             97.8
                          LP                                              90/90                                            88/90
                                                                        (95.9, 100)                                     (92.3, 99.4)
                                                                            100                                              100
                          MP                                               90/90                                           90/90
                                                                        (95.9, 100)                                      (95.9, 100)
a For   the True Negative (TN) category, the reported agreement indicates the percent of negative results.

                     Table 15: Quantitative Site-to-site Reproducibility Results for BD MAX Enteric Viral Panel-NR
                                                              Norovirus                                  Rotavirus                       SPC
    PCR Metric               Parameter
                                                      LP                     MP                   LP                  MP                  TN
                                 N                    90                     90                   88                   90                 90
                                Mean                 29.8                   29.6                 32.3                 31.3               26.8
        Ct. Score
                                 SD                  0.41                   0.43                 0.66                 0.52               0.27
                                %CV                   1.4                    1.4                  2.0                  1.7                1.0
                                  N                   90                     90                   88                   90                 90
                                Mean               4,553.4                4,129.3               737.5                925.2             8,581.7
        Cycle EP
                                 SD                1,269.47               1,769.10              359.91               382.36            1,161.23
                                %CV                  27.9                   42.8                 48.8                 41.3               13.5

Lot-to-lot reproducibility study was performed at one (1) site using three (3) reagent lots. Two (2) operators performed 2 runs per
day, over five (5) distinct days (consecutive or not), for a total of 30 runs. The panels used were the same as described under
the Precision heading, above. Results from 5 days of the accuracy and precision study were used to comprise data for one lot of
reagents for the Lot-to-lot study.
The overall Lot-to-lot reproducibility percent agreements were 99.7 to 100% for TN, and ranged from 97.8 to 100% and
98.9 to 100% and 100% for the LP and MP respectively. The quantitative results across lots and by target are presented in
Tables 16. Ct.Score and the Cycle EP, an internal criteria used to determine a final assay result, was selected as a means
of assessing quantitative assay reproducibility. Mean Ct.Score and the mean Cycle EP values with variance components
(SD and % CV) are shown in Table 17.

                               Table 16: Lot-to-lot Reproducibility Results for BD MAX Enteric Viral Panel-NR
                                                                                     Agreement with Expected Results
                      Category                                          Norovirus                                        Rotavirus
                                                                         (95% CI)                                        (95% CI)
                                                                            100                                              99.7
                         TNa                                              360/360                                          359/360
                                                                        (98.9, 100)                                      (98.4, 100)
                                                                            100                                              100
                          LP                                               90/90                                            90/90
                                                                        (95.9, 100)                                      (95.9, 100)
                                                                            100                                              100
                          MP                                               90/90                                            90/90
                                                                        (95.9, 100)                                      (95.9, 100)
a For   the True Negative (TN) category, the reported agreement indicates the percent of negative results.

                                                                            15
Table 17: Quantitative Lot-to-lot Reproducibility Results for BD MAX Enteric Viral Panel-NR
                                                             Norovirus                                  Rotavirus                           SPC
    PCR Metric             Parameter
                                                     LP                    MP                    LP                    MP                    TN
                              N                      90                     90                  90                     90                    90
                             Mean                   29.7                   29.5                32.0                  31.1                  26.8
     Ct. Score
                              SD                    0.41                   0.35                0.46                  0.35                  0.39
                             %CV                     1.4                    1.2                 1.5                   1.1                   1.5
                               N                     90                     90                  90                     90                    90
                             Mean                 4,945.6                4,948.5              1,117.1               1,345.6               8,993.5
      Cycle EP
                              SD                  2,117.85               2,255.71             267.15                220.59                966.88
                             %CV                    42.8                   45.6                23.9                   16.4                  10.8

Carry-Over / Cross-Contamination
A study was conducted to investigate within-run carryover and between-run carryover while processing samples with high viral load
of analytes in the BD MAX Enteric Viral Panel-NR. A panel made of one high positive member containing one target organism and
one negative member was used to prepare numerous samples. The negative member did not contain any target analyte. Twelve (12)
replicates of the high positive panel member and 12 replicates of the negative panel member were tested in each run by alternating
negative and positive samples. Two (2) operators performed 3 consecutive runs across 3 BD MAX instruments for a total of 9 runs
containing 24 samples. Of the 108 negative samples tested in this study, two samples produced a positive result.
REFERENCES
1. CDC: Estimates of Foodborne Illness in the United States, Located at:
    http://www.cdc.gov/foodborneburden/2011-foodborne- estimates.html
2. Kosek et al. Bulletin of the World Health Organization (2003) 81:197–204.
3. NIH: Bacterial Gastroenteritis, Located at: http://www.nlm.nih.gov/medlineplus/ency/article/000254.htm
4. Petri WA, Miller M, Binder HJ, Levine MM, Dillingham R, and Guerrant RL. Enteric infections, diarrhea, and their impact on
    function and development. J. Clin. Invest. (2008) 118:1277–1290
5. Summary of Notifiable Diseases. available at 
6. CIFOR Analysis of State Legal Authorities. available at 
7. Clinical and Laboratory Standards Institute. Protection of laboratory workers from occupationally acquired infections; Approved
    Guideline. Document M29 (Refer to the latest edition).
8. Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in microbiological and biomedical
    laboratories. Chosewood L.C. and Wislon D.E. (eds) (2009). HHS Publication No. (CDC) 21–1112.
9. BD MAX System User’s Manual (refer to the latest version) BD Life Sciences, Sparks, MD 21152 USA.
10. Clinical and Laboratory Standards Institute. Molecular Diagnostic Methods for Infectious Diseases; Approved Guideline,
    document MM3 (Refer to the latest edition).
11. Clinical and Laboratory Standards Institute. User Protocol for Evaluation of Qualitative Test performance; Approved Guideline,
    Document EP12 (Refer to the latest edition).
12. Catherine Yen, et al. Detection of fecal shedding of Rotavirus vaccine in infants following their first dose of pentavalent Rotavirus
    vaccine. Vaccine 2011 May 31:29(24) 4151–4155

This product is sold under license, and purchase of this product does not include rights to use for certain blood and tissue screening applications, nor
for certain industrial applications.
The purchase of this product allows the purchaser to use it for amplification and detection of nucleic acid sequences for providing human in vitro
diagnostics. No general patent or other license of any kind other than this specific right of use from purchase is granted hereby.
This product is subject to an agreement between Life Technologies Corporation and Becton, Dickinson and Company, and the manufacture, use,
sale or import of this product may be subject to one or more of U.S. patents, pending applications and corresponding foreign equivalents, owned
by Life Technologies Corp. The purchase of this product conveys to the buyer the nontransferable right to use the purchased amount of the product
and components of the product, for use in assays for nucleic acid amplification or detection for the purpose of identifying microorganisms in human
diagnostics. The buyer cannot use this product or its components for manufacturing or for therapeutic or prophylactic use, or sell or otherwise transfer
this product or its components to any third party, or use for any use other than use in human diagnostics. For information on purchasing a license to
this product for purposes other than use in human diagnostics, contact Life Technologies Corporation, Out-Licensing, 5791 Van Allen Way, Carlsbad,
CA 92008, USA, email: outlicensing@lifetech.com.

Technical Information: In the United States contact BD Technical Service and Support at 1.800.638.8663 or www.bd.com.

                                                                           16
You can also read
Next slide ... Cancel