小鼠抗狂犬病毒IgG抗体ELISA检测方法的建立

Establishment of an ELISA assay for the detection of mouse IgG antibody of rabies virus

目的 建立小鼠抗狂犬病毒Ig G抗体ELISA检测方法,用于狂犬病小鼠模型的检测和分析。方法通过正交实验确定样品最佳稀释度和二抗最佳浓度等工作条件;并对该方法的特异性、灵敏性、稳定性等进行分析;与商品化试剂盒一同用于小鼠样品的检测,确定该ELISA方法的符合率。结果 该ELISA方法的样品最佳稀释度是1∶100,二抗最佳浓度是40 ng/m L,阳性临界值为0. 121。该ELISA方法与小鼠常见病毒阳性血清无交叉反应;检测浓度下限为129μg/m L;样本三次重复的变异系数小于10%。与商品化试剂盒的符合率为100%。结论成功建立小鼠抗狂犬病毒Ig G抗体ELISA检测方法,可应用于狂犬病小鼠模型的分析以及疫苗效价的评估工作中。

Objective To establish an ELISA assay for the detection of mouse IgG antibody to rabies virus. Methods Orthogonal experiments were used to determine the optimal dilution of the sample and optimal concentration of the secondary antibody. The stability, specificity and sensitivity test of the assay was determined. The assay result of these samples were compared with those of a commercial kit. Results The optimal dilution of the sample was 1∶100, the optimal concentration of the secondary antibody was 40 ng/ mL, and the cutoff value was 0. 121. There was no crossover with the common mouse viruses, and the minimum detection sensitivity was 129 μg/ mL, and the coefficient variation of the three replicates of the sample was less than 10%. The coincidence rate with the commercial kit was 100%. Conclusions An ELISA assay for the detection of a mouse IgG antibody to rabies virus was successfully established, which can be used for detection and analysis of rabies mouse models.