Standardization of NGS procedure to determine HIV-1 drug resistance mutation to apply in treatment and surveillance of HIV drug resistance

Abstract

In order to determine drug resistance mutations by advanced technology of next - generation sequencing (NGS), we conducted the study to (1) Develop and evaluate the characteristics of NGS procedure to determine HIV-1 drug resistance mutations on pol gene and (2) Apply NGS to detect drug resistance mutations on pol gene in the clinical samples of the patients collected in the North Vietnam. The results showed that the detection limit of the assay was 1000 copies/ml; the accuracy was good with high inter- and intra-repeatability of sequences and mutations; all the major drug resistance mutations detected by Sanger sequencing were determinined by NGS. In application of NGS on the clinical samples, with detection threshold of 5%, in addition to the mutations detected by Sanger sequencing, NGS could detect other major mutations with frequencies from 1.74 - 99.17%. The study outcomes increased the capacity of our laboratory in establishment of the testings based on the advanced technologies, in order to apply in diagnosis, research and surveillance of HIV drug resistance.