Detection of Drug-resistant Mutations in HIV-1 RT Gene Using Magnetic Beads Associated DNA Isolation Technology
磁珠分离DNA技术检测HIV-1逆转录酶基因耐药性突变

In recent years, the emergence of drug-resistant RT gene mutations in HIV-1 strains has become the major reason for AIDS treatment failure. The detection of drug-resistant mutations is important for the treatment choice. In this work, a novel assay for the detection of HIV-1 RT drug-resistant mutations was developed. Drug-resistant and wild strains of HIV-1 B subtype were investigated based on Two-Step MS-PCR. Magnetic beads associated DNA isolation technology and ELISA were employed in this assay to detect point mutations including Y181C and T215F. As a result, this novel assay gave a good P/N ratio and as less as about 5% of mutant type template could be detected, indicating that this assay is highly sensitive and specific. The assay can be completed in relatively less time consumption and applied in high-throughput screening. We suggest that it will be used in clinic assay for the detection of HIV-1 drug-resistant mutations as well as other point mutations.