Defer C, Agut H, Garbarg-Chenon A, Moncany M, Morinet F, Vignon D, Mariotti M, Lefrère J J
Centre Régional de Transfusion Sanguine, Lille, France.
AIDS. 1992 Jul;6(7):659-63. doi: 10.1097/00002030-199207000-00007.
Seven French laboratories tested the specificity and sensitivity of the polymerase chain reaction (PCR) for the detection of HIV-1 DNA.
Following its own PCR protocols, each laboratory independently tested blind two panels of 20 coded peripheral blood mononuclear cell samples collected from HIV-1-seropositive individuals and from HIV-1-seronegative individuals at high or low risk of HIV infection. For the first panel, laboratories were free to select type and number of primers; for the second, all were required to use the two primer pairs Pol 3/4 and MMy 9/10' (Nef 1).
False-positive and false-negative results were observed in all laboratories (concordance with serology ranged from 40 to 100%). In addition, the number of positive PCR results did not differ significantly between high- and low-risk seronegatives. The use of crude cell lysates in DNA preparation produced the same PCR results as phenol-extracted DNA. Discrepancies between laboratories indicated that factors other than primer pairs contributed strongly to laboratory variability.
Our results emphasize the importance of both positive and negative controls in PCR and demonstrate the value of multicentre PCR quality control.
七个法国实验室检测了聚合酶链反应（PCR）检测HIV-1 DNA的特异性和敏感性。
各实验室按照自己的PCR方案，独立对两组共20份编码外周血单个核细胞样本进行盲测，这些样本分别采自HIV-1血清阳性个体以及处于HIV感染高风险或低风险的HIV-1血清阴性个体。对于第一组样本，实验室可自由选择引物的类型和数量；对于第二组，所有实验室都必须使用两对引物Pol 3/4和MMy 9/10'（Nef 1）。
所有实验室均观察到假阳性和假阴性结果（与血清学结果的一致性为40%至100%）。此外，高风险和低风险血清阴性个体的PCR阳性结果数量无显著差异。在DNA制备中使用粗细胞裂解物产生的PCR结果与酚提取的DNA相同。实验室之间的差异表明，除引物对外，其他因素对实验室变异性有很大影响。
我们的结果强调了PCR中阳性和阴性对照的重要性，并证明了多中心PCR质量控制的价值。
