关键词: Alu-PCR, 整合型HIV-1, CD4⁺T细胞

Abstract: Objective To optimize Alu-long terminal repeat(LTR) polymerase chain reaction(PCR) approach to detect integrated HIV-1. Methods The genomic DNA from purified CD4⁺T cells of 20 HIV-1 patients and 10 healthy control subjects was obtained. A pair of primers were designed according to the characteristics of human genome and HIV-1 genome. The first PCR was carried out by using the nested 5' primers from conserved sequences of human Alu and 3' primer from conserved HIV-1 long terminal repeat(LTR) sequences. The product of the first PCR was representative of both cellular DNA upstream of the integration site and integrated HIV-1 LTR. An aliquot of the diluted first PCR product was further subjected to the second round of PCR by using nested HIV-1 LTR-specific primers and then detect the DNA sequence from the product of Alu-LTR PCR. Results After optimized Alu-LTR PCR, integrated HIV-1 was detected in each of the 20 cases of HIV/AIDS. Conclusion After optimization, the sensitivity of Alu-LTR PCR for detection of integrated HIV-1 was markedly improved.

Key words: Alu-polymerase chain reaction, integrate human immunodeficiency virus-1, CD4⁺T cell