Sutthent R, Foongladda S, Likanonskul S, Tunsupasawasdeekul S, Roongpisuthipong A, Chearskul S, Raktham S, Louisirirotchanakul S, Wasi C
Department of Microbiology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
J Med Assoc Thai. 1996 Mar;79(3):142-8.
Nested polymerase chain reaction (nested PCR) was used to separately amplify part of gag, pol, and env genes of human immunodeficiency virus type 1 (HIV-1) to evaluate that primer specific to either gag (SK380/390&SK38/39), pol (JA17/18&JA19/20), or env (JA9/10&JA11/12) genes is suitable for HIV-1 PCR based diagnosis in Thailand. The positive PCR results in 70 HIV-1 infected adults are 100, 97, 89 per cent and in 75 HIV-1 infected infants are 100, 94, 74 per cent by gag, pol, env primer, respectively. The specificity of all three primer sets is 100 per cent. The unamplified samples by pol and env primers were identified as HIV-1 subtype E by PELISA method. False negative in HIV-1 PCR based diagnosis caused by high genetic variation of HIV-1 can be overcome by using several primer sets as shown in this study.
采用巢式聚合酶链反应(nested PCR)分别扩增1型人类免疫缺陷病毒(HIV-1)的gag、pol和env基因部分片段,以评估针对gag(SK380/390&SK38/39)、pol(JA17/18&JA19/20)或env(JA9/10&JA11/12)基因的引物是否适用于泰国基于HIV-1 PCR的诊断。gag、pol、env引物检测70例HIV-1感染成人的阳性PCR结果分别为100%、97%、89%,检测75例HIV-1感染婴儿的阳性PCR结果分别为100%、94%、74%。所有三组引物的特异性均为100%。pol和env引物未扩增的样本通过PELISA方法鉴定为HIV-1 E亚型。如本研究所示,使用多种引物组可克服因HIV-1高度基因变异导致的基于HIV-1 PCR诊断的假阴性。