检测HCV RNA的PCR试剂质控试行参考品的建立

Establishment of A Tentative Quality Control Reference Preparation for PCR Reagent for Detection of HCV RNA

选慢性丙型肝炎病人中HCVRNA含量不等的阳性血清作为阳性标准，采用选自HCV基因5’端非编码区的不同引物来扩增病毒基因，扩增产物与分子量标准进行比较，部分产物用酶切初步分析进行确证。选国家HCV抗体诊断试剂盒临床验证参考品中的阴性样品作为阴性标准，经反复检测显示HCVRNA为阴性。选强阳性血清经5%小牛血清稀释作为灵敏度标准，检测范围为10－5～10－7。应用该参考品对北医肝病研究所的三批HCVRNA诊断试剂盒进行检测，阳性和阴性标准均符合要求，灵敏度为10-5～10-6。

HCV RNA positive reference preparation was established by screened positive serum at differenct level of HCV RNA from patients with chronic hepatitis C. Different primers derived from5' non-coded region of HCV gene was used for amplification of virus gene. Amplified products were compared with molecular weight standards, then a proportion of them were tentatively confirmed by restriction enzyme technique. The negative samples of clinical test by using national HCV antibody diagnostic kit were selected serving as negative standard and repeatedly confirmed to be HCV RNA negative. Strong positive serum diluted with 5% calf serum was selected as sensitivity standard with a sensitivity ranging frorn 10-5~10-5. 3 lots of HCV RNA diagnostic kits (manufactured by Hepatic Disease Institute, Beijing Medical University) were evaluated by the quality control reference preparation. The result showed that both the positive and negative standards of the reference preparation fit in with related requirements, and the sensitivity of it was 10-5~10-6.