应用循环逆转录PCR技术检测丙型肝炎病毒RNA

Detection of Hepatitis C Virus RNA by Circulatory Reverse Transcription-PCR

循环逆转录 (circulatoryreversetranscription ,CRT)是线性增长逆转录cDNA产量的一种新技术。为了将该技术用于检测HCVRNA ,通过改变CRT的循环次数 ,结合竞争PCR ,作出标准曲线。采用 16次CRT加 34次循环PCR检测了 136例HCVELISA阳性、5 4例HCVELISA阴性和 10 8例临床可疑病人全血标本 ,并与逆转录PCR(RT -PCR)和巢式PCR (NT -PCR)的检测结果相比较 ,显示HCVRNA总检出率分别为 :5 7 9%、35 6 %、5 8 6 %。结果提示 ,CRT -PCR是一种简便有效的提高RNA检测灵敏度的方法 。

Circulatory reverse transcription (CRT) reaction is a new technique which can linearly increase cDNA products of reverse transcription reaction and is applied to detect Hepatitis C virus (HCV) RNA. By varying the cycles of CRT, along with the competitive PCR method, we have produced a standard curve. 136 HCV-positive blood samples, 54 HCV-negative blood samples (detected by ELISA) and 108 clinical blood samples were detected with 16 cycles of CRT plus 34 cycles of PCR and a comparative analysis with RT-PCR and NT-PCR detection was conducted. The HCV RNA totally positive rates were 57.9%, 35.6% and 58.6% respectively. The result indicates that the CRT-PCR is a convenient device to enhance the sensitivity of RNA detection and can be applied in the quantification of RNA.