原料血浆混样HCV/HIV-1核酸检测的方法学研究

Establishment of the method of nucleic acids amplification test for HCV/HZV-1 ofpooled source plasma

目的 建立原料血浆混浆核酸检测方法。方法 以国产HCV和HIV核酸扩增 (PCR)荧光定量检测试剂进行WHO标准品的灵敏度、重现性和精密度实验 ,并对 19196份国内原料血浆进行HCVRNA和HIV 1RNA核酸扩增分析。结果 扩增系统能够确保高拷贝数 (2 0 0IU/ml)标准品核酸的检出率 ,对于 <10 0IU/ml的低拷贝数标准品核酸检出率逐渐降低 ;受检原料血浆样本没有检出HCVRNA和HIV 1RNA阳性。结论 核酸扩增方法适用于原料血浆病毒筛查。

Objective To construct the method of nucleic acids amplification test(NAT) for transfusion-transmitted viruses of source plasma.Methods Domestic fluorescence PCR kits for HCV RNA and HIV1 RNA amplification were implemented to assay of sensitivity,reproducibility and robustness of WHO international standards,and then 19196 ELISA negative plasma samples collected from four manufacturers of plasma derivatived were tested for HIV RNA and HCV RNA by NAT.Results The amplification system which had been constructed could assure the detection limits for high copies (200 IU/ml) of nucleic acids from WHO international standard ,and the detection limits for lower than 100 IU/ml of nucleic acids gradually reduced.None of HCV-RNA and HIV/1-RNA was detected in these samples.Conclusion The system of nucleic acids amplification test could be used fuo screening of transfusion-transmitted viruses in source plasma