乙肝表面抗原吖啶酯化学发光定量免疫分析方法的建立
|
摘要
目的建立一种定量检测乙肝表面抗原(HBsAg)的化学发光免疫分析方法并评估其性能。方法利用双抗体夹心法实验原理,用吖啶酯标记多抗,用生物素标记两株单抗,通过磁颗粒链霉亲和素-生物素系统分离固相和液相完成检测。结果本方法的检测限为0.05 IU/mL,线性范围为0.05～150 IU/mL,可溯源至国际标准品,批内变异小于5%,总变异小于8%,浓度高达106IU/mL的强阳样本未出现HOOK效应;5套阳转盘对雅培Architect、西门子centaur和强生VITROS发光试剂相对敏感系数分别是3、0、-3;国家参考盘检出达标;与雅培Architect比较,2000例临床样本检测灵敏度为99.77%,特异性为100%,一致性达99.95%。结论本研究建立的HBsAg定量检测试剂,各项指标均满足临床检测的要求,灵敏度高,特异性好,适合临床推广应用。
Objective To establish a quantitative detection method for Hepatitis B virus surface antigen(HBsAg)and evaluate its performance.Methods According to the principle of double antibody sandwich,anti-HBsAg polyclonal antibody was linked with acridinium ester,and the other two strains of anti-HBsAg monoclonal antibody were linked with biotin,and magnetic beads were coated by streptavidin.The detection was carried out via streptavidin-biotin separation system and washing process.Results The limit of the detection was 0.05 IU/mL,and the linear range was 0.05-150 IU/mL,also the intra-batch and total variation was lower than 5% and 8%,respectively.No HOOK effect was observed in strong positive samples with a concentration of 10~6 IU/mL.The relative sensitivity coefficients of the 5 seroconvertion panel,compared with Abbott Architect,Siemens Centaur and Johnson VITROS chemiluminescent reagents,were 3,0 and-3,respectively.National reference plate detection results conform to the requirements.Compared with Abbott Architect HBsAg kit,the sensitivity and specificity of the 2000 clinical samples were 99.77%,100%,respectively,and the consistency was 99.95%.Conclusion The HBsAg quantitative test reagent established in this study meets the requirements of the clinical application,with high sensitivity and specificity,and is suitable for clinical application.
Objective To establish a quantitative detection method for Hepatitis B virus surface antigen(HBsAg)and evaluate its performance.Methods According to the principle of double antibody sandwich,anti-HBsAg polyclonal antibody was linked with acridinium ester,and the other two strains of anti-HBsAg monoclonal antibody were linked with biotin,and magnetic beads were coated by streptavidin.The detection was carried out via streptavidin-biotin separation system and washing process.Results The limit of the detection was 0.05 IU/mL,and the linear range was 0.05-150 IU/mL,also the intra-batch and total variation was lower than 5% and 8%,respectively.No HOOK effect was observed in strong positive samples with a concentration of 10~6 IU/mL.The relative sensitivity coefficients of the 5 seroconvertion panel,compared with Abbott Architect,Siemens Centaur and Johnson VITROS chemiluminescent reagents,were 3,0 and-3,respectively.National reference plate detection results conform to the requirements.Compared with Abbott Architect HBsAg kit,the sensitivity and specificity of the 2000 clinical samples were 99.77%,100%,respectively,and the consistency was 99.95%.Conclusion The HBsAg quantitative test reagent established in this study meets the requirements of the clinical application,with high sensitivity and specificity,and is suitable for clinical application.
引文
[1]世界卫生组织.慢性乙型肝炎病毒感染预防、关怀和治疗指南[J].中国病毒病杂志,2015,5(5):342-346.
[2]SCHAEFER S.Hepatitis B virus taxonomy and hepatitis B virus genotypes[J],World J Gastroenterol,2007,13(1):14-21.
[3]SCHEIBLAUER H,EL-NAGEH M,DIAZ S.Performance evaluation of 70 hepatitis B virus(HBV)surfaceantigen(HBsAg)assays from around the world by a geographically diverse panel with an array of HBV genotypes and HBsAg subtypes[J].Vox Sang,2010,98(3Pt2):403-414.
[4]国家质量监督检验检疫总局,国家标准化委员会.体外诊断医疗器械生物源性样品中量的测量校准品和质控物质赋值的计量学溯源性[S].中华人民共和国国家标准:GB/T21415-2008/ISO 17511:2003.
[5]王佑春.艾滋病实验室检测技术与质量保证[M].北京:科学出版社,2009:453.
[6]蒋玲丽.国产化学发光法诊断系统检测乙肝表面抗原的评价[D].上海:复旦大学,2009:57-60.
[7]李金明.临床酶免疫测定技术[M].北京:人民军医出版社,2005:100-101.
[8]ZACHER B J,MORICONI F,BOWDEN S.Multicenter evaluation of the elecsys hepatitis B surface antigen quantitative assay[J].Clin Vaccine Immunol,2011,18(11):1943-1950.
[9]候金林,魏来.慢性乙型肝炎防治指南(2015年版)[J].泸州医学院学报,2016,39(1):1-20.
[10]万漠彬,翁心华.干扰素治疗慢性乙型肝炎专家建议[J].中华传染病杂志,2010,28(4):193-200.
[11]HADZIYANNIS E,HADZIYANNIS SJ.Hepatitis B surface antigen quantification in chronic hepatitis B and its clinical utility[J].Expert Rev Gastroenterol Hepatol,2014,8(2):185-195.
[2]SCHAEFER S.Hepatitis B virus taxonomy and hepatitis B virus genotypes[J],World J Gastroenterol,2007,13(1):14-21.
[3]SCHEIBLAUER H,EL-NAGEH M,DIAZ S.Performance evaluation of 70 hepatitis B virus(HBV)surfaceantigen(HBsAg)assays from around the world by a geographically diverse panel with an array of HBV genotypes and HBsAg subtypes[J].Vox Sang,2010,98(3Pt2):403-414.
[4]国家质量监督检验检疫总局,国家标准化委员会.体外诊断医疗器械生物源性样品中量的测量校准品和质控物质赋值的计量学溯源性[S].中华人民共和国国家标准:GB/T21415-2008/ISO 17511:2003.
[5]王佑春.艾滋病实验室检测技术与质量保证[M].北京:科学出版社,2009:453.
[6]蒋玲丽.国产化学发光法诊断系统检测乙肝表面抗原的评价[D].上海:复旦大学,2009:57-60.
[7]李金明.临床酶免疫测定技术[M].北京:人民军医出版社,2005:100-101.
[8]ZACHER B J,MORICONI F,BOWDEN S.Multicenter evaluation of the elecsys hepatitis B surface antigen quantitative assay[J].Clin Vaccine Immunol,2011,18(11):1943-1950.
[9]候金林,魏来.慢性乙型肝炎防治指南(2015年版)[J].泸州医学院学报,2016,39(1):1-20.
[10]万漠彬,翁心华.干扰素治疗慢性乙型肝炎专家建议[J].中华传染病杂志,2010,28(4):193-200.
[11]HADZIYANNIS E,HADZIYANNIS SJ.Hepatitis B surface antigen quantification in chronic hepatitis B and its clinical utility[J].Expert Rev Gastroenterol Hepatol,2014,8(2):185-195.