核酸检测技术在血液筛查中的应用研究

目的大样本、多方法调查深圳地区无偿献血人群中乙肝、丙肝和艾滋病病毒血清学阴性者的核酸阳性率,探讨在我国血液筛查中引进核酸扩增技术的必要性,了解和分析献血者血清学阴性核酸阳性感染状况。方法采用大样本数调查,应用ROCHE PCR-ELISA、PCR-微流芯片、实时荧光PCR方法和CHIRON TMA（转录依赖的扩增技术）多种方法对血清学检测阴性的献血者进行HBV DNA、HCV RNA和HIV-1RNA检测,对乙肝阳性献血者追踪检测ALT和乙肝两对半标志物,对丙肝核酸阳性献血者追踪检测ALT及抗-HCV及HBV DNA和HCV RNA病毒载量。结果共对141288人份血样进行了检测,检出HBsAg（-）、HBV DNA阳性28例,总阳性率为0.020%,其中21例为anti-HBc阳性,占0.015%。HIV-1RNA未检出阳性,17例HBsAg（-）、HBV DNA阳性样本追踪发现,9例发生了血清转换现象,4例呈窗口期特征,所有追踪的HBV DNA阳性献血者ALT检测结果正常。1例anti-HCV（-）、HCV RNA阳性献血者追踪发现为典型窗口期献血,ALT显著升高。结论应采用高灵敏度的核酸扩增技术筛查血液中的乙肝和丙肝病毒,可提高血液安全。

Application on nucleic acid amplification testin blood screening

Objective To investigate and analyze the seronegative and NAT-positive donors,and evaluate the feasibility of using NAT in blood screening. Methods Roche PCR, PCR-CHIP, real time fluorescence PCR and TMA were used for the analysis of HBV DNA, HCV RNA and HIV-1 RNA, respectively, from seronegative samples. HBV positive donors were also analyzed for the serological markers by quantitative PCR. Results 28 out of 141 288 donations were found HBV DNA positive and the HBV DNA positive rate was 0.020%. 21 cases of donations were found anti-HBc positive and the rate was 0.015%. 17 donors were found HBsAg（-） and 9 follow up samples showed seroconversion; 4 donors showed the characteristic of window period. One case was confirmed HCV RNA positive and the donation was in the window period. Conclusion It is necessary to implement the highly sensitive HBV and HCV NAT test for blood screening.