小鼠血清乙型肝炎病毒前S1抗体间接ELISA检测方法的建立和验证

目的  建立小鼠血清乙型肝炎病毒（hepatitis B virus，HBV）前S1抗体的间接ELISA检测方法。方法  以合成的HBV前S1（21-47位氨基酸）多肽包被酶标板，利用方阵滴定法确定间接ELISA方法的最适工作条件，对该法的精密度、特异性和灵敏度进行验证，并与商品化试剂盒进行比较。结果  确定最佳抗原包被浓度为1.0 mg/L，血清稀释度为1︰10。该方法的特异性、灵敏度和精密度均符合检测要求。本法与商品化前S1抗体检测试剂盒测定结果的符合率为98.0%。结论  成功建立了HBV前S1抗体间接ELISA方法，可用于小鼠血清中前S1抗体的检测。

Development and validation of an indirect ELISA for detection of hepatitis B virus PreS1 antibody in mouse serum

Objective To develop an indirect enzyme-linked immunosorbent assay （ELISA） for detection of hepatitis B virus （HBV） PreS1 antibody in mouse serum.Methods Microtiter wells were coated with synthesized HBV PreS1 polypeptides （21-47amino acid） and optimal reaction conditions were determined by criss-cross serial dilution analysis.Validation tests were performed to ensure feasibility of the developed method.Specificity,sensitivity and precision of the method were compared with those of a commercial kit.Results The optimal antigen concentration for coating was 1.0 mg/L and the dilution of serum was 1∶ 10.The specificity,sensitivity,and precision of the method were feasible for quality tests of vaccines.The overall coincidence rate of results by the developed method and the commercial kit was 98.0％ in 150 mouse sera.Conclusion The indirect ELISA suitable for detection of HBV PreS1 antibody in mouse serum is well developed and validated.