HIV-1 GENOTYPING ASSAY FOR GLOBAL SURVEILLANCE OF HIV-1 DRUG RESISTANCE

摘要

Provided herein are new methods, primers, and kits for genotyping HIV-1, including group M viral strains. The methods can be used for HIV-1 drug resistance surveillance and monitoring, for example in resource-poor countries. The disclosed methods can detected more mixed HIV-1 population than previous methods. Given the high efficiency in genotyping diverse HIV-1 group M viral strains from plasma and dried blood spot (DBS) samples and substantial reagent cost saving, the disclosed methods can be used for HIV-1 drug resistance genotyping in both antiretroviral therapy (ART)-naive and -experienced populations for surveillance purposes and patient monitoring.

主权项

1. A method of genotyping HIV-1 in a sample, comprising
contacting a sample comprising HIV-1 DNA obtained from peripheral blood mononuclear cells (PBMC) with a forward nucleic acid primer comprising SEQ ID NO: 1, 2 or 5 and a reverse nucleic acid primer comprising SEQ ID NO: 4 or 6, thereby generating a reaction mixture; incubating the reaction mixture under conditions sufficient to amplify a portion of HIV-1 pol comprising HIV-1 reverse transcriptase (RT) and HIV-1 protease (PR) in the reaction mixture, thereby generating an amplification product; and sequencing the amplification product to determine the genotype of HIV-1 or subjecting the amplification product to allele-specific analysis to determine the genotype of HIV-1.