Study of HIV-1 subtypes in serodiscordant couples attending an integrated counselling and testing centre in Mumbai using heteroduplex mobility analysis and DNA sequencing.

Aims: To determine the prevalent subtypes of HIV-1 in serodiscordant couples. Setting: Integrated Counselling and Testing Centre (ICTC), Department of Microbiology. Study Design: Prospective pilot study. Participants: Thirty HIV-1 serodiscordant couples. Inclusion Criteria: a) Documentation of HIV-1 infection in one partner and seronegative status in the other, current history of continued unprotected sexual activity within the partnership, demonstration that they have been in a partnership for at least 1 year and are not currently on highly active antiretroviral therapy HAART; b) willingness of both partners to provide written informed consent including consent to continued couple counselling for 3 months. Materials and Methods: HIV-1 subtyping was carried out by heteroduplex mobility analysis (HMA) by amplifying env region; and DNA sequencing by amplifying gag region. Results: HIV-1 env gene was amplified successfully in 10/30 samples; gag gene, in 25/30 samples; and both env and gag gene were amplified successfully in 5/30 samples. HIV-1 subtype C was detected from 21 samples; subtype B, from 7; and subtype A, from 2. Sample from 1 positive partner was detected as subtype C by env HMA and subtype B by gag sequencing. Conclusion: HIV-1 subtype C was found to be the predominant subtype of HIV-1 in serodiscordant couples attending our ICTC, followed by HIV-1 subtype B and HIV-1 subtype A, respectively. DNA sequencing was found to be the most reliable method for determining the subtypes of HIV-1.

Quasispecies of HCV before and after Interferon Therapy and Its Dynamic Change Detected by HMA / 中华医院感染学杂志

OBJECTIVE To investigate the quasispecies diversity and the dynamics of quasispecies evolution in patients with chronic Hepatitis C virus(HCV) infection.METHODS Changes of HCV quasispecies before and after interferon(IFN) therapy were detected by heteroduplex mobility analysis(HMA) and by automatic sequencer.RESULTS We found that no changes in viral complexity were observed with therapy,but there were marked changes in viral divergence.The proportion of major pretreatment variants was different in HVR1 in all patients.During the treatment,HCV quasispecies changed significantly.CONCLUSIONS HMA is a simple,rapid and reliable method for detecting HCV quasispecies.Responsiveness to interferon may be related to the changes in viral divergence,but not related to the numbers of viral variants.