全自动血液核酸筛查及阳性献血员追踪的研究

An entirely automatic system of nucleic acid amplification testing in blood screening and follow up study of positive donors

目的建立献血者全自动核酸检测方法,探讨在我国血液筛查中引进全自动核酸筛查方法的可行性。方法在酶联免疫吸附实验(ELISA)筛查血液基础上,选用全自动汇集仪进行血样汇集(24人份),在全自动核酸提取仪上提取样本核酸,应用聚合酶链反应(PCR),在COBASAMPLICOR进行扩增和检测结果,用国际标准核酸质控品考评检出限量,对阳性献血者追踪检测。结果经考评及常规应用表明,全自动汇集、全自动核酸提取及扩增和检测95%的检出限量HBVDNA、HCVRNA和HIV-1RNA分别为38·9、17·4IU/ml和20·6拷贝/ml,95%的可信限分别为[21,323]、[10·5,342]和[12,300]。通过对16512个样本共688汇集池分析,HBVDNA阳性8例,阳性率为0·048%,其中7例为Anti-HBc阳性,其余1例亦转换为阳性。HCVRNA和HIV-1RNA未检出阳性,6例HBVDNA阳性样本追踪发现,3例发生了血清转换现象。结论本实验结果认为全自动血样汇集,全自动核酸提取扩增和检测方法可应用于血液的筛查工作。

Objective To establish the entirely automatic method of nucleic acid amplification testing （NAT） for blood screening and to study the feasibility of NAT. Methods Using entirely automated extraction method to extract nucleic acid , amplified and detected by Roche COBAS AMPLICOR system, evaluating the sensitivity and efficacy. Results The 95% limits of HBV DNA, HCV RNA/HIV-1 RNA tests by automation system were 38.9,16.4IU/ml and 20. 4 copies/ml, 95% Confidence Intervals were [ 21, 323 ], [ 10. 5,342] and [ 12,300] respectively. 8 of 16 512 donations were PCR for HBV DNA positive ,the DNA positive rate was 0. 048%. 7/8 donations were Anti-HBc positive, The last one was also converted positive. No positive HCV RNA and HIV RNA was detected. 3/6 following up samples seroconverted. Conclusions The entirely automatic system can be applied in blood screening.