丙肝病毒抗体化学发光酶免疫分析方法的建立和应用

Establishment and Application of Chemiluminescence Enzyme Immunoassay for Hepatitis C Virus Antibody

【目的】建立丙肝病毒抗体化学发光酶免疫分析方法（CUA）并应用于临床检测。【方法】以辣根过氧化物酶（HRP）为酶标记物，以鲁米诺为发光底物，采用双抗原夹心法检测血样中的抗丙肝病毒（HCV），建立抗HCV化学发光酶免疫分析方法；并与常规使用的抗HCV酶联免疫分析方法（ELISA）同时检测500例血浆标本中的抗HCV，以荧光定量PCR为金标准，探讨其临床应用价值。【结果]CLIA方法敏感度为0．2ng／mL，变异系数〈10％，稳定性好。CLIA和ELISA同时检测500例血浆标本，ELISA检测出7例阳性，CLIA检测出8例阳性，经PCR检测最终确定结果与CLIA相符。【结论]CLIA法检测HCV敏感、准确、稳定性好，适合临床推广应用。

[Objective] To establish the method of chemiluminescence enzyme immunoassay for hepatitis C virus （HCV） antibody and its application in clinical laboratory. [Methods]The double-antigen-sandwich chemiluminescence immunoassay（CLIA） with horseradish peroxidase（HRP） as the enzyme labeling and luminol as the luminescence substrate was established and used for the detection of HCV in 500 blood samples. At the same time, routine antbHCV EIdSA was used to detect the anti-HCV in 500 blood samples. PCR was selected as the gold standard. The clinical values were discussed. [Results] The sensitivity of CLIA was 0. 2 ng/mL and the coefficient of variation was 10%. It had good stability. Seven cases were positive in 500 blood samples detected by ELISA and 8 cases were positive in 500 blood samples detected by CLIS. The final results confirmed by PCR were coincident with those by CLIA. [Conclusion]CLIA is sensitive, precise and stable, and suitable for the extensive application in clinics.