ELISA联合NAT技术在献血者血液筛查和输血残余风险分析中的应用

Application of ELISA combined with nucleic acid testing for blood screening and residual risk analysis in blood donors

目的分析乙型肝炎病毒（hepatitis B virus, HBV）（简称乙肝）表面抗原（HBsAg）酶联免疫法（ELISA）检测阴性献血者血液输血存在乙肝病毒（HBV）感染的残余风险，评价其感染状况。方法选择初筛合格的45 551份无偿献血者样本，采用2种不同的ELISA试剂盒进行HBsAg检测，同时三联荧光病毒单人份核酸扩增检测技术（NAT）检测HBV、丙型肝炎病毒和人类免疫缺陷病毒-1，并对有反应的样本再进行核酸鉴别试验来检测病毒类型，对HBsAg-ELlSA阴性且NAT单反应（HBsAg-/HBV DNA＋）样本进行HBV定量和血清学检测。结果共检出44份HBsAg-/HBV DNA＋样本，42份为隐匿性HBV感染（OBI），2份为窗口期感染（WP）。OBI发生率0.90‰，32份OBI样本HBV DNA定量小于20 IU/ml，OBI检出率不同性别、年龄及献血次数人群间差异有统计学意义（P〈0.05）。血清学检测OBI样本存在6种模式，HBcAb阳性最多39份92.9%（39/42），其中HBcAb阳性样本中76.9%（30/39）HBV DNA定量小于20 IU/ml；29.5%（13/42）的OBI献血者HBeAb和HBcAb同时阳性，其中84.6%（11/13）HBV DNA定量小于20 IU/ml。 结论HBsAg-献血者部分NAT单反应献血者为HBV感染者，多以OBI为主，且HBV DNA浓度低，屏蔽NAT单反应样本对预防经输血传播HBV具有重要意义。

ObjectiveTo analyze the residual risk of transfusion transmitted hepatitis B virus （HBV） infection by enzyme-linked immunosorbent assay （ELISA） method in hepatitis B surface antigen （HBsAg） negative blood donors, and to assess the infection status.MethodsA total of 45551 samples were collected from blood donors.All samples were tested by 2 different ELISA kids of HBsAg and nucleic acid testing （NAT） individually of hepatitis B virus （HBV）, hepatitis C virus （HCV） and human immunodeficiency virus （HIV）. Those ELISA HBsAg negative and NAT single reactive （HBsAg-/HBV DNA＋ ） specimens were analyzed by quantitative detection of HBV DNA and by serologic testing of HBV antigen and antibody.ResultsA total of 44 HBsAg-/HBV DNA＋ samples were detected, including 42 occult HBV infections （OBI） and 2 window period infections （WP）. The detection rate of OBI rate was 0.90‰, and 32 samples of OBI sample HBV DNA was less than 20 IU/ml, and the OBI detection rate was significantly different between different genders, ages and blood donation times （P〈0.05）. In the OBI sample, there were 6 serological models, 92.9%（39/42） OBI samples hepatitis B core antibody （HBcAb） positive, and 76.9%（30/39） HBV DNA in HBcAb positive samples were less than 20 IU/ml; 29.5% （13/42） of OBI blood donors hepatitis B e antigen （HBeAb） and HBcAb were also positive, of whom 84.6% （11/13） were HBV DNA quantitatively 〈20 IU/ml.ConclusionsHBV residual risk of transfusion-transmitted infection may occur through HBsAg- and single NAT reactive blood donors, mainly include OBI, and HBV DNA low level. Blocking of single NAT reactive blood donors could reduce transfusion-transmitted HBV infection.