乙型肝炎病毒前S1抗原化学发光免疫检测方法的建立及初步临床应用

目的建立乙肝病毒前S1抗原（PreSl-Ag）化学发光免疫检测方法，并分析其临床应用价值。方法采用PreSlAg单抗包被微孔板，以辣根过氧化物酶标记的抗乙肝病毒表面抗原的抗体为二抗，结合鲁米诺化学发光底物系统，建立PreSlAg的化学发光免疫检测方法。研制PreSl-Ag诊断试剂企业参考品，并用于分析所建立方法的特异性、灵敏度、稳定性和精密性等试剂性能。所建立方法与市售的PreSl-Ag诊断试剂盒同时比对检测临床血清样本126份，比较检测结果。结果检测结果表明试剂性能符合企业参考品质量标准。批内变异系数6．5％-8．1％、批间变异系数13．2％；试剂盒置37℃考核3d，其稳定性良好。临床样本比对检测结果表明，两种方法相关性良好（阴性符合率96．0％、阳性符合率100％，总符合率为97．6％，Kappa值为0．951，一致性强度为最强）。结论成功建讧了快速、特异、敏感和稳定的PreSlAg化学发光免疫检测方法，适合临床检验推广应用。

Establishment and preliminary clinical application of a chemiluminescence immunoassay for detection of HBV PreSl anti- gen

Objective In order to establish a chemiluminescence immunoassay （CLIA） for detection of HBV PreS1 antigen （PreS1-Ag） and evaluate its clinical application. Methods A CLIA for detection of PreS1 Ag was established by using micro plates coated with a PreSl-Ag mAb, combined with a HRP conjugated anti HBsAg antibody,as well as luminol chemiluminescence substrate system. A manufacturer＇s reference standard were prepared and used for evaluation of specificity,sensitivity,stability and curacy of the established CLIA. PreS1-Ag in sera from 126 clinical samples was also parallel measured by the established method and a reference method,a commercially available ELISA kit. Results The results show that the efficiency of the CLIA reagents met the requirements of manufacturer＇s reference standard. The intra coefficient variations （CV） were among 6. 5%-8. 1% and inter CV 13.2%. The established CLIA showed good correlation with the reference ELISA method （the negative concordance rate,positive concordance rate and overall concordance rate were 96.0%,100% and 97.6%,respectively）,and a highest degree of consistency was observed （the Kappa value=0. 951）. Conclusion So, we have succeeded in establishing a rapid, specific, sensitive and stable CLIA method for detection of PreS1 Ag,which will be fit for widely application in clinical laboratory.