实时荧光PCR结合血浆池策略提高输血安全的研究

Research on transfusion safety using real-time fluorescent PCR combined with minipool

目的:探讨适合于实时荧光PCR技术筛查我国献血者和受血者的血浆池的大小,筛查HBV和HCV免疫学检测阴性的受血者和献血者血液标本,以期发现肝炎病毒血清转换前窗口期的血液标本。方法:用实时荧光定量PCR检测不同拷贝数的HBV DNA和HCV RNA;检测2006年3月~2007年3月期间协和医院接受输血且乙肝三系和丙肝检测结果为阴性的受血者标本,同时检测2006年4月~2006年7月期间孝感市血液中心乙肝三系和丙肝检测结果为阴性的无偿献血者。结果:不同拷贝数的阳性混合标本在1∶24稀释时,HBV DNA和HCV RNA的检测均为阳性;当滴度大于等于1∶48时,则出现了假阴性结果。2006年3月~2007年3月期间协和医院2142名接受输血且所有ELISA检测结果为阴性的患者进行核酸检测,HBV DNA荧光定量测定有2份标本阳性,HCV RNA荧光定量检测没有发现阳性标本;2006年4月~2006年7月期间孝感市血液中心1121名ELISA检测HBV和HCV结果为阴性的无偿献血者进行核酸检测,没有发现阳性标本。结论:采用24份标本混合组成一个检测池进行血液筛查,既准确又经济。

Objective:To study the suitable size of the plasma pool of nucleic acid amplification for blood donor and blood receiver in China, and to screen the immunological HBV or HCV negative blood receiver and blood donor samples detected by the mounted mini-pool model, so as to collect the blood samples infected during 'window phase' and confirm safe blood transfusion. Method:The different titers of HBV DNA and HCV RNA were tested by real-time fluorescent PCR. 2142 samples of immunological hepatitis B virus and hepatitis C virus negative receivers detected by nucleic acid amplification were tested from March 2006 to March 2007 in Union Hospital. At the same time, 1121 samples of hepatitis B virus and hepatitis C virus negative blood donors were tested using the same method from April 2006 to July 2006 in Xiaogan Blood Center.Result:In the samples mixed with different titers of positive HBV and HCV, when diluted at 1∶24, the test results of HBV DNA and HCV RNA were both positive, and when the dilution was at or above 1∶48, the test results of the low levels of HBV DNA or HCV RNA were false negative. From March 2006 to March 2007, 2142 blood samples of patients in Union Hospital who received transfusion and whose results were all negative by ELISA were detected by nucleic acid amplification, two of them were detected positive for HBV DNA and none positive for HCV RNA. From April 2006 to July 2006, 1121 samples of blood donors in Xiaogan Blood Center whose results were all negative by ELISA were detected by nucleic acid amplification, and all were detected negative for HBV DNA and HCV RNA. Conclusion:It is both accurate and economical to use a test pool made up of 24 blood samples for blood screening, which can further ensure transfusion safety.