Development, Validation and Clinical Evaluation of a Low Cost In-House HIV-1 Drug Resistance Genotyping Assay for Indian Patients

Acharya, Arpan; Vaniawala, Salil; Shah, Parth; Misra, Rabindra Nath; Wani, Minal; Mukhopadhyaya, Pratap N.
August 2014
PLoS ONE;Aug2014, Vol. 9 Issue 8, p1
Academic Journal
Human Immunodeficiency Virus-1 (HIV-1) drug resistance genotyping assay is a part of clinical management of HIV-1 positive individuals under treatment with highly active antiretroviral therapy (HAART). Routine monitoring of drug resistance mutations in resource limited settings like India is not possible due to high cost of commercial drug resistance assays. In this study we developed an in-house, cost effective HIV-1 drug resistance genotyping assay for Indian patients and validated it against the US-FDA-approved ViroSeq HIV-1 drug resistance testing system. A reference panel of 20 clinical samples was used to develop and validate the assay against ViroSeq HIV-1 drug resistance testing system which was subsequently used to genotype a clinical panel of 225 samples. The Stanford HIV database was used to identify drug resistant mutations. The analytical sensitivity of the assay was 1000 HIV-1 RNA copies/ml of plasma sample while precision and reproducibility was 99.68±0.16% and 99.76±0.18% respectively. One hundred and one drug resistant mutations were detected by the in-house assay compared to 104 by ViroSeq system in the reference panel. The assay had 91.55% success rate in genotyping the clinical panel samples and was able to detect drug resistant mutations related to nucleoside reverse transcriptase inhibitor (NRTI), non-nucleoside reverse-transcriptase inhibitor (NNRTI) as well as protease inhibitor (PI) classes of antiretroviral drugs. It was found to be around 71.9% more cost effective compared to ViroSeq genotyping system. This evaluation of the assay on the clinical panel demonstrates its potential for monitoring clinical HIV-1 drug resistance mutations and population-based surveillance in resource limited settings like India.


Related Articles

  • Regimen selection in the OPTIONS trial of HIV salvage therapy: drug resistance, prior therapy, and race-ethnicity determine the degree of regimen complexity. Tashima, Karen T.; Mollan, Katie R.; Na, Lumine; Gandhi, Rajesh T.; Klingman, Karin L.; Fichtenbaum, Carl J.; Andrade, Adriana; Johnson, Victoria A.; Eron, Joseph J.; Smeaton, Laura; Haubrich, Richard H. // HIV Clinical Trials;Aug2015, Vol. 16 Issue 4, p147 

    Background: Regimen selection for highly treatment-experienced patients is complicated. Methods: Using a web-based utility, study team members reviewed antiretroviral (ARV) history and resistance data and recommended individual ARV regimens and nucleoside reverse transcriptase inhibitor (NRTI)...

  • A Cluster Randomized Trial of Routine HIV-1 Viral Load Monitoring in Zambia: Study Design, Implementation, and Baseline Cohort Characteristics. Koethe, John R.; Westfall, Andrew O.; Luhanga, Dora K.; Clark, Gina M.; Goldman, Jason D.; Mulenga, Priscilla L.; Cantrell, Ronald A.; Chi, Benjamin H.; Zulu, Isaac; Saag, Michael S.; Stringer, Jeffrey S. A. // PLoS Clinical Trials;Mar2010, Vol. 7 Issue 3, Special section p1 

    Background:The benefit of routine HIV-1 viral load (VL) monitoring of patients on antiretroviral therapy (ART) in resource-constrained settings is uncertain because of the high costs associated with the test and the limited treatment options. We designed a cluster randomized controlled trial to...

  • Emergence of HIV-1 drug resistance mutations among antiretroviral-naive HIV-1-infected patients after rapid scaling up of antiretroviral therapy in Thailand.  // Journal of the International AIDS Society;2012, Vol. 15 Issue 1, p12 

    The article focuses on a study which aims at determining the prevalence and associated factors of primary HIV-1 drug resistance in Thailand. It states that an observational study was conducted among the antiretroviral-naïve HIV-1-infected Thai patients. It mentions that there is a significant...

  • Subtype variability, virological response and drug resistance assessed on dried blood spots collected from HIV patients on antiretroviral therapy in Angola. Carolina Garrido; Natalia Zahonero; Diana Fernándes; Dulcelina Serrano; Ana Rita Silva; Nelia Ferraria; Francisco Antúnes; Juan González-Lahoz; Vincent Soriano; Carmen de Mendoza // Journal of Antimicrobial Chemotherapy (JAC);Mar2008, Vol. 61 Issue 3, p694 

    : Background Subtype variability may influence treatment response and selection of drug resistance mutations in HIV-positive patients on antiretroviral therapy. : Patients and methods A retrospective study was performed on specimens collected on dried blood spots (DBS) from HIV-positive...

  • ART treatment interruption may lead to virological failure, drug resistance.  // Infectious Disease News;Sep2012, Vol. 25 Issue 9, p6 

    The article focuses on a study which suggested that HIV patients who resumed antiretroviral therapy (ART) after treatment interruption experienced virological failure and drug resistance after resuming their previous regimen.

  • Reverse transcriptase drug resistance mutations in HIV-1 subtype C infected patients on ART in Karonga District, Malawi.  // AIDS Research & Therapy;2011, Vol. 8, p38 

    The article reports on the use of a consensus sequencing approach for genotyping reverse transcriptase (RT) from HIV-1 subtype C infected individuals on antiretroviral therapy (ART). The results reveal the presence of mutations associated with drug resistance to the therapy. Drug resistance to...

  • Treatment for Adult HIV Infection: 2004 Recommendations of the International AIDS Society-USA Panel. Yeni, Patrick G.; Hammer, Scott M.; Hirsch, Martin S.; Saag, Michael S.; Schechter, Mauro; Carpenter, Charles C. J.; Fischl, Margaret A.; Gatell, Jose M.; Gazzard, Brian G.; Jacobsen, Donna M.; Katzenstein, David A.; Montaner, Julio S. G.; Richman, Douglas D.; Schooley, Robert T.; Thompson, Melanie A.; Vella, Stefano; Volberding, Paul A. // JAMA: Journal of the American Medical Association;7/14/2004, Vol. 292 Issue 2, p251 

    Context Substantial changes in the field of human immunodeficiency virus (HIV) treatment have occurred in the last 2 years, prompting revision of the guidelines for antiretroviral management of adults with established HIV infection. Objective To update recommendations for physicians who provide...

  • Seroconversion Following Nonoccupational Postexposure Prophylaxis against HIV. Roland, Michelle E.; Neilands, Torsten B.; Krone, Melissa R.; Katz, Mitchell H.; Franses, Karena; Grant, Robert M.; Busch, Michael P.; Hecht, Frederick M.; Shacklett, Barbara L.; Kahn, James O.; Bamberger, Joshua D.; Coates, Thomas J.; Chesney, Margaret A.; Martin, Jeffrey N. // Clinical Infectious Diseases;11/15/2005, Vol. 41 Issue 10, p1507 

    Background. The efficacy of antiretroviral postexposure prophylaxis (PEP) against infection with human immunodeficiency virus (HIV) following occupational exposures has prompted the use of PEP after nonoccupational exposures. There are, however, important differences between occupational and...

  • HIV Antiretroviral Postexposure Prophylaxis: A Cautionary Note. Cohen, Myron S.; Kashuba, Angela D. M.; Gay, Cynthia // Clinical Infectious Diseases;11/15/2005, Vol. 41 Issue 10, p1514 

    Presents the authors' opinions on the role of antiretroviral therapy in preventing the sexual transmission of human immunodeficiency virus (HIV) infection. Discussion on clinical trials to evaluate the efficacy of antiretroviral therapy; Views on the benefits of pre-exposure prophylaxis;...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics