Antiretroviral treatment (ART) use in India requires information on baseline drug resistance mutations and polymorphisms in the protease (Pr) and reverse transcriptase (RT) genes of HIV-1 strains from treatment-naïve individuals. We report resistance predictor mutations and polymorphisms in the Pr and the RT sequence of non-clade B HIV-1 strains from ART naïve individuals. The genotypic resistance assay was done on 93 treatment-naïve individuals. The sequences were analysed by Stanford HIV drug resistance data for genotypic drug resistance analysis and REGA HIV-1 subtyping tool. Phylogenetic tree was generated with MEGA 4 for quality control. Ninety-two strains belonged to clade C and one to clade A (A1). Amino acid substitutions were seen at positions associated with drug resistance in Pr gene--10, 24, 74 (each 3%) and position 82 (11%). Substitutions were seen at positions 41 (1%), 100 (1%), 101 (6%), 103 (2%), 179 (6%) and 181 (1%) of the RT sequence known to confer drug resistance in clade B. Polymorphisms in HIV-1 pol gene among treatment-naïve individuals were similar when compared with previous data. One strain each had Y181C substitution, T74S and E35G substitutions in the Pr and one had A98G, K101R and L210FL substitutions in RT.

An alternative technology for the estimation of T cells based on a microcapillary technique (Guava Technologies, Hayward, CA) was compared to FACSCount (Becton Dickinson, San Jose, CA). Samples from 51 human immunodeficiency virus-infected and 21 healthy individuals were tested. The correlation (r) of the two systems for CD4؉ T cells was 0.994, and the coefficient of variation was 6.5%, establishing equable performance between the two technologies.The number of human immunodeficiency virus (HIV)-infected individuals continues to rise in India ever since it was first detected in Vellore, India (6). A high proportion of these infected individuals belong to the economically lower strata of society, and hence it is essential that the supporting health care systems operate in a cost-effective manner. CD4 ϩ T-cell counts are essential for monitoring disease progression and as indicators of when treatment should be started (4). Flow cytometry is the widely used method for analyzing CD4 counts, including in India. More recently a system based on a new technology developed by Guava Technologies (Hayward, CA) has been introduced. This technology has been used only in a small number of centers and requires more widespread testing for user acceptability. In this study, we investigated the performance of the Guava EasyCD4 System in relation to FACSCount.Blood samples were collected between 8:00 a.m. and 10:00 a.m. from 51 HIV-infected individuals who had come to the Clinical Virology department of a tertiary care center in India (south) for their CD4 ϩ /CD8 ϩ T-cell estimation and also from 21 normal healthy individuals from October 2004 through December 2004 after informed consent was obtained in K 2 EDTA vacutainer tubes. Of the 51 samples from HIV-infected individuals, 8 were done in duplicates to investigate the reproducibility of results. In addition to these eight samples, samples from four HIV-infected individuals were tested in quadriplicates. Samples from another seven HIV-infected individuals were stored at room temperature (20 to 25°C) and were retested after 24 h to look for any difference in the count. In order to examine the interpersonnel variation in testing, samples from two healthy individuals were processed and tested by five different laboratory personnel only for CD4 counts.Absolute CD4 ϩ /CD8 ϩ T-cell counts were estimated by the Guava EasyCD4 System, which allows enumeration of CD4 ϩ and/or CD8 ϩ lymphocytes in human peripheral blood. The instrument used two fluorescence parameters in combination with forward scatter (FSC) to identify cells. The reagents consisted of a monoclonal anti-human CD3 antibody conjugated to the tandem dye phycoerythrin (PE)-Cy5 and monoclonal anti-human CD4 and CD8 antibodies conjugated to PE. The antibody staining solutions were used to stain 10-l portions of the blood samples. For each sample, two separate tubes were used: one for determining the absolute CD4 ϩ T-cell count (monoclonal antibody CD3PE-Cy5 and CD4PE were added) and the other for determining the CD8 ϩ T-cell c...

This study showed an association of EBV and CMV load with CD4+ T cell counts, WHO staging and elevated liver enzymes. These viral infections can accelerate HIV disease and multiplex real-time PCR can be used for the early detection. Genotype 1 and 2 of EBV and genotype gB1 and gB2 of CMV were the prevalent in the HIV-1 subtype C-infected south Indians.