核酸检测在血液HBV筛查中的应用研究

Application of NAT detection for HBV in blood screening

目的:了解沈阳地区乙型肝炎病毒表面抗原(HBsAg)阴性献血者中乙型肝炎病毒(hepatitis B virus,HBV)感染状况,探讨HBV核酸扩增检测(nucleic acid amplification testing,NAT)技术应用于血液筛查的意义。方法:在酶联免疫法(enzyme immunoassay,EIA)检测的基础上,应用TaqMan实时荧光聚合酶链式反应(PCR)方法对HBsAg EIA阴性血液标本进行HBV DNA的NAT检测。对NAT阳性标本进一步做乙型肝炎病毒血清标志物(HBV Marker,HBV-M)及核酸定量检测。结果:共检测了105,152例HBsAg阴性的血液标本,检出HBV DNA阳性标本15例,阳性率0.014%。Abbott酶联免疫试剂检测发现,15例标本中有6例标本的HBV-M五项指标检测全部阴性,9例为HBsAg阴性但其它标志物阳性。其中10例HBV DNA阳性标本再经Roche试剂进行核酸定量检测,HBV DNA核酸含量最高为149 IU/ml。结论:在HBsAg ELA阴性献血者中仍有极少数的HBV感染者;核酸扩增检测和酶联免疫检测互补,能够缩短血液筛查中HBV检测窗口期,特别是对提高HBsAg阴性血液标本中HBV感染检出率具有重要价值。

Objective：To investigate the HBV infection status of the HBsAg negative blood donors in Shenyang and explore the significance of nucleic acid amplification testing（NAT） in blood donor screening.Methods：Based on enzyme immunoassay（EIA） detection,the HBsAg EIA negative blood specimens were detected for HBV DNA by TaqMan real-time fluorescence PCR.The NAT positive specimens were then used to detect HBV marker and quantify nucleic acid further.Results：Among 105,152 blood specimens which were negative for HBsAg,15（0.014%） were positive for HBV DNA.When detecting those 15 specimens by using Abbott reagent,there were 6 specimens negative for all the HBV antigen/antibody detection,the other 9 specimens were only negative for HBsAg.Among those specimens,10 was tested,the highest amount of HBV DNA was 149 IU/ml by using Roche reagent.Conclusion：There is still minority of HBV infection among the HBsAg EIA negative blood donors.Combining NAT detection with EIA,the ＂window period＂ of HBV detection could be shortened,which is highly significant for improving the relevant ratio in large-scale blood donors screening.