Current HIV Research

Charles Wood  
University of Nebraska
Lincoln, NE


Pre-Screening HIV-1 Reverse Transcriptase Resistance Mutations in Subtype B Patients Using a Novel Multiplex Primer Extension Assay

Author(s): Martin Roelsgaard Jakobsen, Anni Aggerholm, Louise Bruun Jorgensen, Alex Laursen and Lars Ostergaard

Affiliation: Department of Infectious Diseases, Aarhus University Hospital, Skejby, Brendstrupgaardvej 100, 8200 Aarhus N, Denmark.

Keywords: HIV-1, Drug Resistance, Minority mutation, Genotyping, Primer extension


Antiretroviral therapy is standard treatment for HIV-infected patients. Such therapy has decreased mortality and morbidity, but treatment success is often jeopardized by the emergence of viral drug resistance. Moreover, in recent years there has been a reported rise in the incidence of transmitted drug resistance, highlighting the importance of pretreatment resistance screening. In this report, we describe the development and utility of a sensitive multiplex approach for detecting mutations conferring drug resistance to HIV-1 reverse transcriptase inhibitors. This protocol, termed HIVSNaPshot, utilizes a multiplex primer extension assay with capillary electrophoresis reporting altered nucleotides at nine important drug resistance mutation positions. Mutations were successfully detected to levels of 5% in viral quasispecies populations. Furthermore, although developed and optimised for HIV-1 subtype B, drug resistance mutations could also be detected in most non-B subtypes. Comparison of the HIV-SNaPshot with the commercial Viroseq genotyping system in 10 patients gave similar results, but importantly, additional resistance mutations were identified in several patients by the HIV-SNaPshot assay. Thus, the HIV-SNaPshot is a method capable to support standard genotyping for the determination of minority HIV-1 resistance mutations, with equivalent and perhaps greater sensitivity than Viroseq.

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Article Details

Page: [398 - 409]
Pages: 12
DOI: 10.2174/157016209788680615