HPV Detection and genotyping in males from the city of Córdoba, Argentina
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184 Revista Argentina de Microbiología (2010) 42:0325-7541
ISSN 184-188
INFORME BREVE Revista Argentina de Microbiología (2010) 42: 184-188
HPV Detection and genotyping in males from the city of
Córdoba, Argentina
F. VENEZUELA1*, L .E. KREMER2, X. KIGUEN1, C. CUFFINI1
1
Instituto de Virología “Dr.J. M. Vanella” U.N.C. Calle Enfermera Gordillo Gómez s/n (entre avenidas Enrique Barros y
Valparaiso) Ciudad Universitaria, Córdoba (5016); 2Hospital Nacional de Clínicas-Córdoba.
Calle Santa Rosa 1500, B° Alberdi, Córdoba (5000)
*Correspondence. Email: venezuelafernando@hotmail.com
ABSTRACT
A wide range of human papillomavirus (HPV) types can infect the anogenital region of males. Although there is a vast
knowledge on HPV infections in women as well as on their association with cervical cancer, the study of HPV infections
in males is scarce and controversial. The aim of the present work was to detect and typify HPV infections of the
anogenital region in males and analyze the associated risk factors in the population studied. Anogenital samples from
37 patients (30 of whom were HIV carriers) attending the Infectology Service at the Hospital Nacional de Clínicas in
Córdoba, Argentina, were studied. Nine of these patients tested HPV-positive and five out of these nine were found to
have mixed infections, being 18 and 61 the most frequent genotypes. There was a significant correlation between the
HPV-positive patients and those having an HPV-compatible lesion or AIDS. The present work is the first study in the
city of Cordoba which contributes relevant results to the knowledge of HPV infection and to the possible implementation
of measures for its prevention.
Key words: Male, genotyping, HPV, AIDS, anogenital, Córdoba
RESUMEN
Detección y genotipificación de VPH en varones de la ciudad de Córdoba, Argentina. Un amplio espectro de
tipos de virus papiloma humano (VPH) puede infectar la zona anogenital de los varones. Si bien existe un vasto
conocimiento de la infección por VPH en las mujeres y su asociación con el cáncer de cérvix, el estudio de la infección
por VPH en los varones ha sido escaso y sus resultados controvertidos. El presente trabajo tuvo como objetivo
detectar y tipificar infecciones por VPH en la región anogenital de varones y analizar los factores de riesgo asociados
en la población estudiada. Se estudiaron muestras anogenitales de 37 pacientes (30 portadores del VIH) que asistie-
ron al Servicio de Infectología del Hospital Nacional de Clínicas de la ciudad de Córdoba. Nueve resultaron positivas
para VPH, de las que 5 correspondían a infecciones mixtas. Los genotipos de mayor frecuencia fueron el 18 y el 61.
Hubo una correlación significativa entre los resultados VPH positivos en aquellos pacientes con lesión compatible o
con sida. Nuestro trabajo es el primero de su tipo realizado en la ciudad de Córdoba y aporta resultados relevantes
para el conocimiento de la infección por VPH y la implementación de medidas de prevención.
Palabras clave: Varón, genotipificación, HPV, sida, anogenital
Human papillomavirus (HPV) infection has become anogenital region, b) their last sexual partner presented
one of the most important sexually transmitted diseases a lesion compatible with HPV in the anogenital region,
(13, 14). HPV causes not only benign lesions (like and c) more than three sexual partners in the previous
condylomata acuminata), but also a wide range of year. The average age of patients was 36.9 years (range
preneoplastic lesions and carcinoma. Although the rela- 21-57 years).
tionship between HPV and the etiopathogeny of cervical The Infectology Service provides care to patients with
cancer is well known, there are few studies on HPV in- different types of lesions. However, it is interesting to point
fection in male anogenital lesions. In addition, it has been out that over 80% of the patients included in this study
proposed that males play an important role as reservoirs were carriers of the human immunodeficiency virus
and transmitting agents of HPV infection (4, 11). (twenty-nine of HIV and one of HIV-HCV). In accordance
During March-June 2009, in order to detect and typify with the Center for Disease Control and Prevention (CDC),
HPV infections in the anogenital region of males, we stud- 16 of the 29 HIV carriers were classified as HIV-AIDS
ied 40 samples from 37 patients attending the Infectology (A3: 3; C1: 2; C2: 6 and C3: 5).
Service at the Hospital Nacional de Clínicas in Córdoba, An epidemiological chart was performed for each pa-
Argentina, who presented at least one of the following tient and, according to the data collected, either a urethral
conditions: a) a lesion compatible with HPV in the swab/penile brushing or a brushing of the anal/perianalHPV Detection and genotyping in males from the city of Córdoba, Argentina 185
region was obtained, since these samples are consid- This could be due to the fact that the primers used in this
ered optimal to evaluate the infection in the anogenital study detected the mucosal HPV types (1). This may in-
region (5). A total of 28 urethral swabs/penile brushing dicate that the lesions were caused by non-mucosal HPV
and 12 anal/perianal were obtained. In three of the pa- types, since the β-globin gene was the one amplified in
tients reporting both passive and active sexual inter- all the samples. Another hypothesis is that the lesions
course, both types of samples were collected. The rea- could have been caused by another etiologic agent, since
sons why each patient attended the Infectology Service they were classified as HPV-compatible, always under a
are detailed in Table 1. clinical criterion.
Of the 29 patients that attended the Infectology The largest number of patients presenting lesions was
Service for reasons other than an HPV-compatible lesion, observed in the compatible range of 21 to 30 years of
six presented this kind of lesion in the anogenital region age (57%), which correlates with the positive results for
and 12 stated that they had had previous HPV lesions HPV (28.6%).
(Table1). Several articles have reported the detection of HPV
Of the 14 patients who presented an HPV-compatible in men, whose infection rates range between 1.0% and
lesion, two presented perianal condyloma acuminata and 82.9% (3) depending on the population studied, the
12 presented flat warts (four in the perianal region, three anatomic site of the sample, the collection method and
in the glans penis and five on the surface of the penis). the sensitivity of the technique for the detection of HPV
The samples were collected in 500 µl phosphate-buffered (6, 7, 9).
saline (PBS). DNA was then extracted using the com- In the present work, the percentage of patients who
mercial AccuPrep Genomic DNA Extraction Kit (Bioneer attended the Infectology Service presenting an HPV in-
Inc., CA, USA), according to the manufacturer’s instruc- fection was 24.4%, a value comparable to the 33% found
tions. A 450-bp segment, corresponding to the L1 region in the males attending a private hospital specialized in
of the viral genome, was amplified by PCR, using the sexually transmitted diseases in the USA (15). It is worth
degenerate primers MY09 and MY11 (1). The product was pointing out that the methodology used, the population
detected by electrophoresis in agarose gel at 1.5% using included and the type of sample obtained were similar in
an U.V. transilluminator. both studies.
The Beta globin gene was used as a DNA preserva- Table 2 shows that HPV-positive patients, with more
tion marker. Viral genotypification was carried out by than three sexual partners in the year previous to our
analysis of restriction fragment length polymorphism study, reported not having used a condom (never / some-
(RFLP), following the technique described by Bernard et times) in any of their sexual encounters. If we point out
al. (1). A statistical analysis was carried out with Epi Info™, that all of them had at least a high school degree, we
Version 3.5.1, 2008 (http://www.cdc.gov/epiinfo/). could conclude that the educational background does not
Of all the samples analyzed, nine were positive for guarantee the knowledge and application of the meas-
HPV and belonged to patients that presented an HPV- ures to prevent a sexually-transmitted infection.
compatible lesion (Table 3). In addition, five patients that This table also shows that three out of the six patients
presented an HPV-compatible lesion were HPV-negative. presenting an HPV-compatible lesion who had attended
the Infectology Service for other reasons tested positive.
It is interesting to point out that one of them (patient No.
Table 1. Distribution according to the inclusion conditions (n = 37)
4) presented a mixed infection, whose genotypes be-
longed to the high risk group. This patient had had sexual
Reason for consultation
intercourse with the opposite sex, which indicates the lack
of knowledge of the population of the risks of this infec-
HPV-compatible lesions 8
tion, not only for themselves, but also for their sexual
Clinical check up and in search of medication 22
partner(s). This result is very important for the purposes
Other (traumatisms, allergies, pneumonia, etc.) 7
of determining the guidelines for the prevention of HPV
infection.
Reason for enrollment in the study
Table 2 also indicates that of all the genotypes identi-
fied (6, 11, 16, 18, 26, 44, 45, 59, 61 and 68), the most
HPV-compatible lesion (A) 6
frequent ones were 18 and 61, and shows that in pa-
Sexual partner with HPV-compatible lesion (B) 3
tients with condylomata acuminata, only the low-risk HPV
More than three sexual partners in the last year (C) 17
genotypes 61 and 11 were detected.
A+B 2
Of the three patients from whom both kinds of sam-
A+C 5
ples were taken, only one (patient n° 6) was positive
B+C 3
(genotype 18-68) and corresponded to the sample taken
A+B+C 1
from the perianal region.186
Table 2. Characteristics of HPV-positive patients
HPV-positive 1 2 3 4 5 6 7 8 9
patients
Age 23 31 25 30 45 52 30 37 47
(1)
Reason for A+B A+B A A+C A+C A A+B+C A A
enrolment in the
study
Reason for Compatible Compatible Clinical Other Compatible Compatible Compatible Clinical Compatible
consultation lesion lesion check up lesión lesion lesion check up lesion
(2)
Type of sample 1 1 1 1 2 2 2 1 2
Characteristics of Flat wart Flat wart Flat wart Flat wart Condyloma Flat wart Flat wart Flat wart Condyloma
the lesion acuminata acuminata
Lesion location Glans Glans Penis surface Penis surface Perianus Perianus Perianus Penis surface Perianus
Time of the lesion 2 months 2 months 1 month 2 months 6 months 1 month 2 months 0.5 months 12months
Sexual preference Heterosexual Heterosexual Homosexual Heterosexual Homosexual Homosexual Homo sexual Homosexual Homosexual
No. of patners 1 >3 1 >5 >5 1 >10 1 1
Use of condom Always Never Always Never Sometimes Always Never Always Sometimes
Educational Finished Finished Finished Finished Finished Finished Finished Finished Did not finish
background high school high school tertiary level high school tertiary level university tertiary level high school university
education education education
Classification of — — Yes Yes Yes Yes Yes Yes Yes
HIV carrier (CDC) [A3] [C2] [C3] [C2] [C2] [C3]
Genotype 44 61-18 6-61 16-45 61 18-68 18 26-59 11
(1)
Reason for enrollment in the study: (A) Lesion compatible with HPV, (B) sexual partner with lesion compatible with HPV, (C) more than three sexual partners. (2)Type of sample: (1) urethral swab/penile brushing,
(2) anal/perianal brushing
Revista Argentina de Microbiología (2010) 42: 184-188HPV Detection and genotyping in males from the city of Córdoba, Argentina 187
Regarding the genotype frequency, no significant dif- precautions had been taken in the Infectology Service
ferences between the low-risk and the high-risk HPV types where this study was carried out.
were observed. It can be noticed that every two HPV- The RFLP analysis of three (HIV positive) out of the
positive patients with compatible lesions, one could have seven HPV positive-patients, showed DNA bands that
at least one infection caused by a high-risk genotype. were not attributed to other genotypes present. Since a
Likewise, 80% of the patients with an AIDS diagnosis by significant association with the presence of multiple HPV
the presence of the disease marker (C2 and C3) were infections in HIV patients has been reported (8), this
positive for at least one high-risk genotype, although for result suggests that some of them may have had co-infections
HIV-carriers any genotype should be considered high risk. with other HPV genotypes. Importantly, the two HCV-
It should also be noted that, since the implementation of positive patients were negative for HPV detection.
antiretroviral therapy has failed to reduce the incidence HPV detection was significantly associated with two
of some cancers caused by HPV (10), all the necessary factors: AIDS and the HPV-compatible lesion, which is in
Table 3. Frequency of detection of HPV vs risk factors
Total HPV - (%) HPV + (%) p
Total N° of patients 37 28 (75.6) 9 (24.4)
HIV: YES 30 23 (76.7) 7 (23.3) 0.77110
AIDS 0.04980
Yes 16 10 (62.5) 6 (37.5)
No 14 13 (92.9) 1 (7.1)
HIV: NO 7 5 (71.4) 2 (28.6)
Sexual preference 0.56580
Homosexual 24 19 (79.0) 5 (21.0)
Heterosexual 10 7 (70.0) 3 (30.0)
Bisexual 3 2 (66.7) 1 (33.3)
LESION 0.00001
Yes 14 5 (35.7) 9 (64.3)
No 23 23 (100.0) 0 (0.0)
Partner with lesion 0.46890
Yes 9 6 (66.7) 3 (33.3)
No 28 22 (78.6) 6 (21.4)
Use of condom 0.23210
Always 17 13 (76.5) 4 (23.5)
Never 6 3 (50.0) 3 (50.0)
Sometimes 14 12 (85.7) 2 (14.3)
N° of partners 0.28000
1 11 6 (54.5) 5 (45.5)
>3 10 9 (90.0) 1 (10.0)
>5 4 3 (75.0) 1 (25.0)
>10 8 6 (75.0) 2 (25.0)
>20 4 4 (100.0) 0 (0.0)
Educational background 0.66800
Did not finish elementary school 1 1 (100.0) 0 (0.0)
Finished elementary school 3 3 (100.0) 0 (0.0)
Did not finish high school 2 2 (100.0) 0 (0.0)
Finished high school 13 9 (69.2) 4 (30.8)
Did not finish tertiary level education 1 1 (100.0) 0 (0.0)
Finished tertiary level education 6 3 (50.0) 3 (50.0)
Did not finish university 5 4 (80.0) 1 (20.0)
Finished university 6 5 (83.3) 1 (16.7)188 Revista Argentina de Microbiología (2010) 42: 184-188
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Acknowledgements: We thank all the patients enrolled in
1107-14.
this study and Drs Fanny N. Cohen, Carlos M. Quinteros and
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Luis Allende, from the Infectology Service at the Hospital Nacio-
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nal de Clínicas in Córdoba, Argentina, for their invaluable
11. Partridge JM, and Koutsky LA. Genital human papillo-
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12. Sirera G, Videla S, Herranz P, Corzo-Delgado J. Human
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