Human immunodeficiency virus type 1 (HIV-1) genotyping in Rio de Janeiro, Brazil: assessing subtype and drug-resistance associated mutations in HIV-1 infected individuals failing highly active antiretroviral therapy.

In order to assess the human immunodeficiency virus type 1 (HIV-1) drug resistance mutation profiles and evaluate the distribution of the genetic subtypes in the state of Rio de Janeiro, Brazil, blood samples from 547 HIV-1 infected patients failing antiretroviral (ARV) therapy, were collected during the years 2002 and 2003 to perform the viral resistance genotyping at the Renageno Laboratory from Rio de Janeiro (Oswaldo Cruz Foundation). Viral resistance genotyping was performed using ViroSeq Genotyping System (Celera Diagnostic-Abbott, US). The HIV-1 subtyping based on polymerase (pol) gene sequences (protease and reverse transcriptase-RT regions) was as follows: subtype B (91.2%), subtype F (4.9%), and B/F viral recombinant forms (3.3%). The subtype C was identified in two patients (0.4%) and the recombinant CRF_02/AG virus was found infecting one patient (0.2%). The HIV-1 genotyping profile associated to the reverse transcriptase inhibitors has shown a high frequency of the M184V mutation followed by the timidine-associated mutations. The K103N mutation was the most prevalent to the non-nucleoside RT inhibitor and the resistance associated to protease inhibitor showed the minor mutations L63P, L10F/R, and A71V as the more prevalent. A large proportion of subtype B was observed in HIV-1 treated patients from Rio de Janeiro. In addition, we have identified the circulation of drug-resistant HIV-1 subtype C and are presenting the first report of the occurrence of an African recombinant CRF_02/AG virus in Rio de Janeiro, Brazil. A clear association between HIV-1 subtypes and protease resistance mutations was observed in this study. The maintenance of resistance genotyping programs for HIV-1 failing patients is important to the management of ARV therapies and to attempt and monitor the HIV-1 subtype prevalence in Brazil.

HIV-1 diversity in Brazil: genetic, biologic, and immunologic characterization of HIV-1 strains in three potential HIV vaccine evaluation sites. Brazilian Network for HIV Isolation and Characterization.

The Brazilian Network for HIV Isolation and Characterization was established for the surveillance of HIV variability in Brazil. Here, we report characterization of HIV strains and virus-specific immune responses from 35 clinical samples collected from three potential HIV vaccine sites. Three genetic subtypes of HIV-1 were identified by heteroduplex mobility assay (HMA) B (in 82.9% of the samples), F (14.3%), and C (2.9%). Phylogenetic analysis based on the C2V3/env DNA sequence from all 25 specimens examined was 100% concordant with HMA results. Four variants of subtype B with different tetrapeptides at the tip of the V3 loop were found: the GPGR motif (North American), GWGR motif (Brazilian B"), and two minor variants, GFGR and GPGS, as previously detected. No significant association was found between HIV-1 subtypes and the mode of transmission or biologic properties of HIV-1 isolates (derived from 88.6% of the specimens). Only 5 of 16 isolates studied were neutralized by the autologous sera. Consistent with previous results, no relation between viral subtype and peptide enzyme-linked immunosorbent assay (ELISA) seroreactivity or neutralization was evident. This study also demonstrated the effectiveness of the collaborative approach followed by Brazilian scientists when addressing a complex subject such as HIV variability.

Validation of national algorithms for the diagnosis of sexually transmitted diseases in Brazil: results from a multicentre study.

OBJECTIVE: To validate STD flow charts for the management of genital discharge and genital ulcer currently recommended by the National STD Control Programme in Brazil. METHODS: A study was conducted in five Brazilian STD clinics from January to June 1995. After an interview, a clinical examination was performed by a physician, who recorded a presumptive diagnosis, based on his/her clinical experience. This diagnosis was compared with a gold standard laboratory diagnosis in order to calculate sensitivity, specificity, and positive predictive value of the clinical diagnosis. The validity of the simulated national flow charts was assessed using the same method. RESULTS: A total of 607 men and 348 women participated in the study. Gonorrhoea was the aetiology most frequently detected in men with urethral discharge. The sensitivity of the clinical diagnosis was far lower than the sensitivity fo the national flow chart, using the syndromic approach, for both gonococcal and chlamydial urethritis. Adding a simple laboratory test (Gram stain) to the national flow chart increased the specificity and positive predictive value for gonorrhoea. Among the women with vaginal discharge, a cervical infection was detected in 17%, a vaginal infection in 74%, and mixed infection in 9%. The sensitivity of the diagnosis for cervical infection increased from 16% (clinical aetiological approach) to 54% (when adding a syndromic approach) and to 68% when adding a risk assessment, as in the national flow charts. The cure or improved rate of genital ulcers was 96% after 1 week. CONCLUSIONS: The results of the study will help to convince policy makers and those involved in training healthcare workers in Brazil of the public health advantages of the syndromic approach, as an essential part of STD/HIV control activities.