Human papillomavirus DNA detection and typing in male urine samples from a high-risk population from Argentina.

The aim of the present study was to evaluate the first void urine (FVU) as a non-invasive sampling method for HPV detection and genotyping in a high-risk population. Men presenting with HPV associated penile lesions or HPV positive partners attending a urological department in La Plata, Argentina were enrolled for HPV detection and genotyping. DNA from 185 first-void urine samples was evaluated for the presence of HPV by nested polymerase chain reaction using MY09/11 and GP05/06 primers. The viral genotype was analyzed by means of the single-stranded conformation polymorphisms (SSCP) method. Seventy-three percent (135/185) of the FVU specimens were positive for HPV-DNA. The viral prevalence in patients with HPV-DNA positive partners was 68.8% (77/112), and 79.5% (58/73) of patients with penile lesions were found to be HPV positive. The most frequent viral type was HPV-11 (26.7%), followed by HPV-6 (23%), HPV-16 (21.5%), HPV-18 (6%), and HPV-31 (4.4%). In this study, 11.1% (15/135) of the HPV positive specimens were double infections. These results indicate that high-risk HPVs can be found in clinical lesions in a high percentage (43.8%), as simple or double infections. In this sense, the male population represents an important reservoir for the virus and may play a role in the transmission and perpetuation of the infection in the general population. The method described below provides a tool for detection and typing of HPV-DNA using samples obtained by non-invasive techniques and thus easy to obtain.

Chlamydia trachomatis and Human papillomavirus infections in cervical disease in Argentine women.

OBJECTIVES: To examine the prevalence of Human papillomavirus and Chlamydia trachomatis DNA in cervical samples among women with normal and abnormal cervical cytology from La Plata, Argentina. METHODS: Two hundred and seventy-nine women (200 with cervical neoplasia or ICC and 79 women with normal cytology) provided cervical samples for the detection of HPV and C. trachomatis DNA by PCR-based assays. RESULTS: HPV DNA increased with the cervical lesion severity, ranging from 30% among women with normal cytology to 99-100% among women with HSIL or ICC. C. trachomatis DNA prevalence increased from low levels in women with normal cytology (11%) to 47% in those with HSIL, but was uncommon among ICC patients (20%). Among women with normal cytology, C. trachomatis prevalence was higher in HPV DNA positive (12.5%) than HPV DNA negative women (10.9%), but this difference was not significant. CONCLUSIONS: HPV prevalence in the general population is slightly higher than those reported for other developing countries. C. trachomatis DNA positivity was associated with a higher risk of both LSIL and HSIL lesions, but not with ICC.

Herpes simplex virus genotyping: multiple optional PCR-based RFLP systems and a non-isotopic single-strand conformation polymorphism method.

Polymerase chain reaction (PCR)-based methods for herpes simplex virus (HSV) types I/II genotyping are described. These methods are based on the single-stranded conformation properties of DNA molecules obtained by PCR (PCR-SSCP) and restriction fragment analysis of PCR products (PCR-RFLP). With the aim to analyse these two genotyping techniques, genomic DNA from the standard viral strains KOS and G, and also 79 normal cervical samples were studied for HSV I/II. Sequence analysis showed 19 different RFLP possible systems for HSV typing. Five systems were used in this study and all led to the expected fragments obtained by sequence analysis. PCR-SSCP showed distinct patterns for both viral types and had 100% of concordance with PCR-RFLP results. The prevalence of herpes simplex virus in the normal cervical samples were 21.5% (17 out of 79), the most frequent viral type was HSV-2, with a prevalence of 71% (12 out of 17). Both techniques appeared suitable for HSV I/II genotyping and are easy to perform in most clinical laboratories.

The c-myc activation in cervical carcinomas and HPV 16 infections.

Despite the prominent role for Human Papillomavirus (HPV) infection in the development of genital cancer, other genetic or environmental co-factors have also been involved. Studies of c-myc activation in cervical carcinomas have reported that gene over-expression (mainly gene amplification) are common in cervical squamous cell carcinomas and may correlate with the biologic behavior of the neoplasm. Using PCR based technology, DNAs from 79 normal cervical samples and 225 abnormal cervical tissue scrapes were analyzed for HPV detection and typing and for c-myc gene amplification. Significant differences were found between the different cyto/histology groups (P<0.0001) and also with HPV high-risk infected samples (P<0.0002). In this sense, we showed that the average c-myc copy number increased according to the histological grade of the lesion (OR=6.3, CI=2.1-18.8). Also, the results showed that the infection with HPV 16 was tightly associated with c-myc amplification (OR=10.6, CI=3.1-36). These results could indicate that oncogene amplification take place in pre-invasive stages of cervical disease and could cooperate not only in tumor progression but also in cell transformation. Moreover, the results strongly associate the c-myc gene amplification to the infection with the oncogenic HPV 16, showing that the pattern of virus infection and oncogene activation could be specific for different viral genotypes.

The p53 codon 72 genotypes in HPV infection and cervical disease.

OBJECTIVE: [corrected] An experimental study has indicated that individuals homozygous for the Arg-encoding allele of p53 gene may have an increased susceptibility to HPV-related cervical cancer but many epidemiological studies have failed to repeat this result. Many epidemiological studies have failed in the attempt to repeat this results. The aim of the present work was to investigate whether the p53 arginine allele confers a risk factor for cervical carcinogenesis. STUDY DESIGN: Using PCR based technology, DNAs from 90 normal cervical samples and 205 abnormal cervical tissue scrapes were analyzed for the type of HPV present and p53 codon 72 polymorphism. RESULTS: Non statistically significant differences were found for the frequencies of p53 genotypes in the different cytological/histological groups (chi2=1.4; P=0.97) nor for the risk for HPV infection (chi2=1; P=0.9). CONCLUSION: This study showed that polymorphism at codon 72 of TP53 gene is not associated with an increased susceptibility to cervical disease and/or HPV infection in the Argentine women population.

Amplificación del proto-oncogén HER-2/neu asociada a la infección por VPH de bajo riesgo en citología exfoliativa cervical / HER-2/neu prot-oncogen amplification related to low risk VPH infection in cervical exfoliative cytology

Se analizaron 69 muestras positivas para el virus papiloma humano (VPH) tipos -6, -16 y -18, y 24 muestras con citología cervical normal negativas para el VPH como grupo control. El análisis de la amplificación génica del proto-oncogén HER-2/neu se realizó utilizando la técnica de coamplificación con locus de referencia. Se encontró una asociación entre la amplificación del gen HER-2/neu y el grupo viral de "bajo riesgo" (VPH-6) (p < 0.005). Dentro de este grupo, se observó una asociación entre la amplificación del proto-oncogén y el status citopatológico CIN I (p < 0.01). Debido a que la mayoría de las muestras CIN I analizadas presentaron un patrón coilocítico, la amplificación de HER-2/neu parecería estar relacionada con este tipo de alteración celular. Por otra parte, sería importante estudiar la amplificación génica y la expresión de HER-2/neu en los diferentes estadios de las neoplasias intraepiteliales cervicales a fin de poder evaluar su papel en la progresión del cáncer cervical.