Objective: To establish a rapid nested multiplex PCR assay for subtyping HIV-1 CRF01_AE, CRF07_BC, CRF08_BC, B, and C strains prevailing in Guangxi.
Method: Subtype-specific primers were designed for these subtypes based on their gag sequences. The subtypes of HIV-1 samples from Guangxi were determined by nested multiplex PCR and DNA sequencing and phylogenetic analysis, respectively, and then the sensitivity and the specificity of nested multiplex PCR were calculated.
Results: Nested multiplex PCR could correctly classify the 5 known-subtype samples, and were not reactive to all HIV-negative samples. Of the 72 HIV-positive samples, 66 were correctly identified as CRF01_AE, CRF07_BC, CRF08_BC, and B by this assay, giving a sensitivity of 91.7% (66/72), and a specificity of 100%.
Conclusion: This assay is a simple, fast, and cost-effective subtyping method for HIV-1 CRF01-AE, CRF07_BC, CRF08_BC, and B strains prevailing in Guangxi.