Eron J J, Gorczyca P, Kaplan J C, D'Aquila R T
Infectious Disease Unit, Massachusetts General Hospital, Charlestown.
Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3241-5. doi: 10.1073/pnas.89.8.3241.
Polymerase chain reaction (PCR) DNA quantitation (PDQ) susceptibility testing rapidly and directly measures nucleoside sensitivity of human immunodeficiency virus type 1 (HIV-1) isolates. PCR is used to quantitate the amount of HIV-1 DNA synthesized after in vitro infection of peripheral blood mononuclear cells. The relative amounts of HIV-1 DNA in cell lysates from cultures maintained at different drug concentrations reflect drug inhibition of virus replication. The results of PDQ susceptibility testing of 2- or 3-day cultures are supported by assays measuring HIV-1 p24 antigen production in supernatants of 7- or 10-day cultures. DNA sequence analyses to identify mutations in the reverse transcriptase gene that cause resistance to 3'-azido-3'-deoxythymidine also support the PDQ results. With the PDQ method, both infectivity titration and susceptibility testing can be performed on supernatants from primary cultures of peripheral blood mononuclear cells. PDQ susceptibility testing should facilitate epidemiologic studies of the clinical significance of drug-resistant HIV-1 isolates.
聚合酶链反应（PCR）DNA定量（PDQ）药敏试验可快速、直接地测定1型人类免疫缺陷病毒（HIV-1）分离株的核苷敏感性。PCR用于定量外周血单核细胞体外感染后合成的HIV-1 DNA的量。在不同药物浓度下培养的细胞裂解物中HIV-1 DNA的相对量反映了药物对病毒复制的抑制作用。通过检测7天或10天培养上清液中HIV-1 p24抗原产生的试验，支持了对2天或3天培养物进行PDQ药敏试验的结果。用于鉴定导致对3'-叠氮-3'-脱氧胸苷耐药的逆转录酶基因突变的DNA序列分析也支持PDQ结果。使用PDQ方法，可对外周血单核细胞原代培养物的上清液进行感染性滴定和药敏试验。PDQ药敏试验应有助于对耐药HIV-1分离株临床意义的流行病学研究。
