Design of a genotyping assay for identifying and monitoring HIV-2 drug resistance
Introduction: Human immunodeficiency virus type 2 (HIV-2) is a causative agent of acquired immunodeficiency syndrome (AIDS) but is distinct from the more prevalent HIV-1. HIV-1 and HIV-2 can rapidly acquire mutations which confer drug resistance (DR). HIV-2 can be treated with some HIV-1 antiretrovirals, but is intrinsically resistant to others. Genotyping assays for HIV-1 are used to identify DR mutations and guide treatment. No clinically approved genotyping assays exist for HIV-2, making treatment difficult. Objectives: Our overall objective is to develop and validate an HIV-2 DR genotyping assay for clinical use. The goal of this project was to develop a set of pol region amplification and sequencing primers and to collect and organize DR mutation information by genomic region with appropriate evidence. Methods: HIV-2 information was gathered from literature and public databases. Amplification and sequencing primers were aligned for compatibility with reference sequences from HIV-2 group A and B using sequence analysis software and tools available from the Los Alamos HIV databases (http://www.hiv.lanl.gov). Results: Primers were developed that successfully amplified and sequenced the pol region of HIV-2 group A viruses. An annotated pol region gene map was created using HIV-2 group A and B reference sequences, which will allow for sequence analysis at positions known to confer DR and for comparison with closely related viruses. Conclusion: We have made significant progress in developing the HIV-2 DR genotyping assay. Our future goals are to optimize the assay, validate it for clinical use and create an HIV-2 rules-based DR interpretation tool.
Chronic disease management and prevention
Clinical medicine applied in public health
Protection of the public in relation to communicable diseases including prevention or control
Public health biology
Describe the process involved in designing an HIV-2 genotyping assay for drug resistance.
Discuss the role of a gene map as an interpretation tool for HIV-2 sequences.
Keyword(s): HIV/AIDS, Infectious Diseases
Presenting author's disclosure statement:
Qualified on the content I am responsible for because: The student was involved in all aspects of the test design process including data collection, identification and organization of HIV-2 specific primers and drug resistance information from the literature, creation of the HIV-2 gene map and selection of the primers for use. The student was not involved in laboratory procedures related to optimizing the primers or the amplification and sequencing reactions. The specific laboratory procedures for the HIV-2 genotype assay will be described elsewhere.
Any relevant financial relationships? No
I agree to comply with the American Public Health Association Conflict of Interest and Commercial Support Guidelines,
and to disclose to the participants any off-label or experimental uses of a commercial product or service discussed
in my presentation.