乙型肝炎病毒外膜大蛋白检测方法的建立

Establishment of a detection method for hepatitis B virus large surface protein （HBV-LP）

目的建立人血清中乙型肝炎病毒外膜大蛋白（HBV-LP）的酶联免疫吸附试验（ELISA）检测方法。方法辣根过氧化物酶标记HBV-LP单抗，优化各类反应液的工作浓度和各种反应条件，建立双抗体夹心的ELISA检测方法；同时评价所建立方法的灵敏度、特异性、稳定性等性能指标。结果所建立方法的灵敏度为5ng／ml，批内、批间变异均小于10％。在4℃和37℃条件下分别进行了3、5、7d的稳定性考察，线性相关系数均〉0．98，标准偏差〈10％。结论成功建立了血清中HBV—LP的检测方法，该方法性能能够满足临床检测的需求。

Objective To establish enzyme-linked immunosorbent assay （ELISA） for detection of hepatitis B virus large surface protein（HBV-LP） in serum. Methods A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of HBV-LP as the catalytic enzyme. Several reactions liquid＇s concentration and reaction conditions were optimized. The method was evaluated in all aspects such as sensitivity, specificity, stability and so on. Results The detection limit was 5ng/ml. Inter- assay and intra-assay RSD were both less than 10%. After stored at 4℃ and 37℃ for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0. 98 and RSD lower than 10%. Conclusion Established ELISA for determination of serum HBV-LP has high sensitivity and repeatability.