E6 and E7 variants of human papillomavirus-16 and -52 in Japan, the Philippines, and Vietnam.

Human papillomavirus (HPV) has several intragenotypic variants with different geographical and ethnic distributions. This study aimed to elucidate the distribution patterns of E6 and E7 (E6/E7) intragenotypic variants of HPV type 16 (HPV-16), which is most common worldwide, and HPV-52, which is common in Asian countries such as Japan, the Philippines, and Vietnam. In previous studies, genomic DNA samples extracted from cervical swabs were collected from female sex workers in these three countries and found to be positive for HPV-16 or HPV-52. Samples were amplified further for their E6/E7 genes using type-specific primers and analyzed genetically. Seventy-nine HPV-16 E6/E7 genes were analyzed successfully and grouped into three lineages: European (Prototype), European (Asian), and African-2. The prevalences of HPV-16 European (Prototype)/European (Asian) lineages were 19.4%/80.6% (n = 31) in Japan, 75.0%/20.8% (n = 24) in the Philippines, and 0%/95.8% (n = 24) in Vietnam. The 109 HPV-52 E6/E7 genes analyzed successfully were grouped into four lineages, A-D; the prevalences of lineages A/B/C/D were, respectively, 5.1%/92.3%/0%/2.6% in Japan (n = 39), 34.4%/62.5%/0%/3.1% in the Philippines (n = 32), and 15.8%/73.7%/7.9%/2.6% in Vietnam (n = 38). The distribution patterns of HPV-16 and HPV-52 lineages in these countries differed significantly (P < 0.000001 and P = 0.0048, respectively). There was no significant relationship between abnormal cervical cytology and either HPV-16 E6/E7 lineages or specific amino acid mutations, such as E6 D25E, E6 L83V, and E7 N29S. Analysis of HPV-16 and HPV-52 E6/E7 genes can be a useful molecular-epidemiological tool to distinguish geographical diffusion routes of these HPV types in Asia.

Prediction of response to treatment of chronic hepatitis B with pegylated interferon in the Philippines.

The response marker for interferon has not been investigated fully for hepatitis B viruses (HBVs) in the Philippines where novel subtypes B5 and C5 were recognized recently. The prediction parameters for interferon treatment were assessed, with emphasis on the mutation patterns in the basal core promoter and precore regions in patients with chronic hepatitis B. Seventeen HBeAg-positive patients were stratified according to response to treatment with pegylated interferon based on HBe seroconversion and HBV load. Intra-patient distributions of wild-type strains (A1762, G1764) and variants (T1762, A1764) were analyzed using HBV-DNA amplification and subsequent molecular cloning. The rate of variants (T1762, A1764) harbored by a patient was higher among responders (41.2% and 31% per person on average) than among non-responders (2.4% and 2.4%) to treatment with pegylated interferon at the baseline, respectively (P < 0.05). The rate of variants (T1762, A1764) harbored by responders (41.2% and 31%) decreased to 1.7% and 1.7%, and wild-type strains (A1762, G1764) conversely became majority (98.3% and 98.3%) after treatment with pegylated interferon, respectively. HBV strains harbored by two of six responders and a patient with lower baseline load (1.0 x 10(4) copies/ml) showed genotype shift from A to other genotypes, where genotype A disappeared preferentially after the loss of HBeAg and genotypes B and C formed a major population. These results suggest that the HBV variants (T1762, A1764) and HBV genotype A in the Philippines have an advantage in the response to pegylated interferon. These results warrant a large-scale examination for further precise prediction of the response to treatment with interferon.

High-risk HPV types in lesions of the uterine cervix of female commercial sex workers in the Philippines.

In order to prevent cervical cancer, vaccines against human papilloma virus types 16 (HPV-16) and 18 (HPV-18) have been implemented worldwide. However, the HPV types that cause cancer can differ according to geographical area and ethnicity. In this new era of the HPV vaccine, it is important to elucidate the prevalent HPV types in each area. Therefore, the prevalence of HPV infection and cervical abnormalities among 369 female commercial sex workers in the Philippines were examined. HPV L1 gene was amplified by polymerase chain reaction (PCR) using modified GP5+/6+ primers, and genotyping was performed by sequencing cloned PCR products. HPV DNA was detected in 211 (57.2%) women, among whom 46 HPV types were identified. HPV-52 was most common and multiple-type infection was observed in 44.5%. Among 56 women with abnormal cervical cytology (low- and high-grade squamous intraepithelial lesions and adenocarcinoma in situ), HPV-52 was most common (23.2%), followed by HPV-16 (19.6%), -58 (10.7%), and -67 (10.7%). Only 27% of these women were positive for HPV-16 and -18. Multivariate analysis revealed that HPV-16, -39, -52, -67, and -82 were significantly associated with abnormal cytology. Repeated analysis of HPV-52 single-positive samples using the original GP5+/6+ PCR primers produced negative results in 57% of cases, suggesting that the prevalence of HPV-52 infection may have been underestimated in previous studies, and the current vaccines may not be sufficient for preventing infection and the development of premalignant lesions of the cervix in women in the Philippines.

A natural inter-genotypic (2b/1b) recombinant of hepatitis C virus in the Philippines.

The prevalence study and the characterization of hepatitis C virus (HCV) was carried out in the Philippines and the sequence determination of the 5'-untranslated region (5'-UTR)-Core and the NS5B regions of HCV was carried out in this study. An HCV strain (SE-03-07-1689) collected in Metro Manila, Philippines, belonged to discordant subtypes, 2b and 1b in 5'-UTR-Core and NS5B regions, respectively. The 9.3 kb sequence of this strain including the entire open reading frame was compared with those of the reference strains retrieved from the HCV sequences database (GenBank/EMBL/DDBJ) and indicated a recombination event. The computation of the sequence similarity mapped a crossover point within the NS3 region. This is the second report on the inter-genotype recombinant of HCV and the third when an intra-genotype recombinant is included. This recombinant strain, SE-03-07-1689, is designated tentatively as RF3_2b/1b according to the suggestions used for the other two HCV recombinants.

Rapid spread of hepatitis C virus among injecting-drug users in the Philippines: Implications for HIV epidemics.

From the trends of human immunodeficiency virus (HIV) epidemics in South and Southeast Asia, it was postulated that an HIV epidemic would start as a blood-borne infection among injecting-drug users in the Philippines. In 2002, 560 individuals were recruited in Metro Cebu, Philippines and tested for HIV, hepatitis C virus (HCV), and hepatitis B virus (HBV) infections. The seroprevalence of anti-HCV among injecting-drug users (70.1%, 61/87) was significantly higher than those among inhalation drug users (16.3%, 7/43; P = 0.00; OR = 12), sex workers (0%, 0/130; P = 0.00; OR = infinity), antenatal clinic attendees (0%, 0/100; P = 0.00; OR = infinity), and students/health care workers (2%, 4/200; P = 0.00; OR = 115). The seroprevalence of HBsAg among injecting-drug users (10.3%, 9/87) was significantly higher than those among sex workers (2.3%, 3/130; P = 0.01; OR = 4.9), and antenatal clinic attendees (3%, 3/100; P = 0.04; OR = 3.7), but was not statistically different from those among inhalation drug users (9.3%, 4/43; P = 0.9) and students/health care workers (4.5%, 9/200; P = 0.06). None of the study population was reactive to anti-HIV antibody. The HCV strains obtained from the injecting-drug users belonged to either genotype 1a or 2b and the strains in each genotype clustered closely to each other. There was no dual infection with genotype 1a and 2b. These results suggest that the HCV infection in injecting-drug users may be emanating rapidly from limited number individuals in Metro Cebu, Philippines.