建立用于检测HIV-1的基于核酸序列扩增的酶联免疫吸附测定法

Development of a NASBA-ELISA based method for detection of HIV-1

目的建立用于检测HIV-1的基于核酸序列扩增的酶联免疫吸附测定(NASBA-ELISA)法。方法根据HIV-1基因组长末端重复序列设计引物对HIV-1RNA进行基于核酸序列扩增(NASBA),结合微孔板中液相杂交和酶标显色反应检测核酸扩增物,并对该检测方法的灵敏度、特异度及其对临床样本的检测结果进行评价。结果该方法可检测到0.1fg/μL的RNA,且对HIV-1具有特异性。在120例临床样本的检测中,其检测准确度高于常规的实时荧光定量反转录聚合酶链反应(fqRTPCR)法。结论 NASBA-ELISA方法能灵敏并特异地检测HIV-1。

Objective To develop a nucleic acid sequence-based amplification （NASBA） coupled ELISA method for detection of human immunodeficiency virus-1 （HIV-1）. Methods The HIV-1 RNA was amplified by NASBA using gene special primers, which were designed according to the long terminal repeat nucleotide sequence of the HIV-1 genome. And the NASBA products were hy bridized into the mieropiates in liquid phase and colorimetric changes were observed. Then the sensibility, specificity and clinical test results of this method were evaluated. Results This method could detect as little as 0. 1 fg/μL RNA,and was highly specific for HIV-1 detection. Results from 120 clinical samples showed its accuracy was higher than traditional real-time RT-PCR. Conclusion The NASBA-ELISA method can detect HIV-1 sensitively and specifically.