Abstract:

Objective To validate laboratory-developed human immunodeficiency virus-1（HIV-1）integrase genotyping sequence assay which can be applied to assess the level of genotyping drug resistance to HIV-1 integrase inhibitors. Methods According to the World Health Organization self-developed gene sequence analysis method validation recommendation，RNA was extracted from 20 HIV-1 positive samples，and the gene fragments of HIV-1 integrase region were amplified and sequenced. The accuracy of laboratory-developed HIV-1 integrase genotyping sequence assay was evaluated by comparison with virus quality assurance（VQA） consensus. Amplification sensitivity was evaluated by amplification success rate. Precision and reproducibility were evaluated by repeated tests of the same sample. Results All the 20 samples had a nucleotide agreement rate of >98% with VQA consensus. The multi-well amplification rate of 10 high viral load samples（>10 000 copies·mL^-1） and 5 low viral load samples （1 000-5 000 copies·mL^-1） was 100%. The results of 5 complex wells of 4 samples of the same batch，as well as 5 assays of 5 samples，met the requirement of nucleotide agreement rate of >98% in 90% sample pairwise comparison. Conclusions The accuracy，amplification sensitivity，precision and reproducibility of this assay meet all the requirements and are suitable for HIV-1 integrase genotyping.

Key words: Human immunodeficiency virus-1, Integrase genotyping sequence assay, Genotyping drug resistance determination