alexa Comparing Peripheral Blood Mononuclear Cell DNA and Cir
ISSN 2155-6113

Journal of AIDS & Clinical Research
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Research Article

Comparing Peripheral Blood Mononuclear Cell DNA and Circulating Plasma viral RNA pol Genotypes of Subtype C HIV-1

Lauren Banks1, Sharareh Gholamin1*, Elizabeth White1, Lynn Zijenah2 and David A. Katzenstein1

1Center for AIDS Research, Stanford University Medical Center, Stanford, CA, USA

2Department of Immunology, University of Zimbabwe, Harare, Zimbabwe

*Corresponding Author:
Sharareh Gholamin, MD
Center for AIDS Research
Stanford University Medical Center
300 Pasteur Drive, Room S-146 Grant Bldg
Stanford, CA 94305, USA
Tel: 650-723-8291
Fax: 650-725-2395
E-mail: [email protected]

Received Date: January 01, 2012; Accepted Date: February 12, 2012; Published Date: February 15, 2012

Citation: Banks L, Gholamin S, White E, Zijenah L, Katzenstein DA (2012) Comparing Peripheral Blood Mononuclear Cell DNA and Circulating Plasma viral RNA pol Genotypes of Subtype C HIV-1. J AIDS Clinic Res 3:141. doi:10.4172/2155-6113.1000141

Copyright: © 2012 Banks L, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

Introduction: Drug resistance mutations (DRM) in viral RNA are important in defining treatment most likely to provide effective antiretroviral therapy (ART) in HIV-1 infected patients. Detection of DRM in peripheral blood mononuclear cell (PBMC) DNA is another source of information, although the clinical significance of DRMs in proviral DNA is less clear.

Materials and Methods: From 25 patients receiving ART at a center in Zimbabwe, 32 blood samples were collected. Dideoxy-sequencing of gag-pol identified subtype and resistance mutations from plasma viral RNA and proviral DNA. Drug resistance was estimated using the calibrated population resistance tool on www.hivdb.stanford. edu database. Numerical resistance scores were calculated for all antiretroviral drugs and for the subjects’ reported regimen. Phylogenetic analysis as maximum likelihood was performed to determine the evolutionary distance between sequences.

Results: Of the 25 patients, 4 patients (2 of which had given 2 blood samples) were not known to be on ART (NA) and had exclusively wild-type virus, 17 had received Protease inhibitors (PI), 18, non-nucleoside reverse transcriptase inhibitors (NNRTI) and 19, two or more nucleoside reverse transcriptase inhibitors (NRTI). Of the 17 with history of PI, 10 had PI mutations, 5 had minor differences between mutations in RNA and DNA. Eighteen samples had NNRTI mutations, six of which demonstrated some discordance between DNA and RNA mutations. Although NRTI resistance mutations were frequently different between analyses, mutations resulted in very similar estimated phenotypes as measured by resistance scores. The numerical resistance scores from RNA and DNA for PIs differed between 2/10, for NNRTIs between 8/18, and for NRTIs between 17/32 pairs. When calculated resistance scores were collapsed, 3 pairs showed discordance between RNA and DNA for at least one PI, 6 were discordant for at least one NNRTI and 11 for at least one NRTI. Regarding phylogenetic evolutionary analysis, all RNA and DNA sequence pairs clustered closely in a maximum likelihood tree.

Conclusion: PBMC DNA could be useful for testing drug resistance in conjunction with plasma RNA where the results of each yielded complementary information about drug resistance. Identification of DRM, archived in proviral DNA, could be used to provide for sustainable public health surveillance among subtype C infected patients.

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