乙型肝炎病毒核心抗体IgM时间分辨荧光免疫分析试剂盒的研制

Development of time-resolved fluoroimmunoassay kit for detection of IgM antibodies against hepatitis B core antigen

目的利用时间分辨荧光免疫分析(TRFIA)技术建立乙型肝炎病毒抗HBc-IgM的检测试剂盒。方法采用捕获法建立抗HBcIgM TRFIA检测试剂盒,对自制试剂盒的各项性能指标进行评估。结果用自制质控品检测分析内和分析间的精密度分别为4.8～7.2%,7.5～8.6%;与抗-HBs,抗-HBcIgG和抗-HBe无交叉反应;对584份血清样本进行检测并与国产酶联免疫法试剂盒对比,结果显示自制试剂盒的灵敏度更高;300份健康者血清样本检测结果表明,该试剂盒的cutoff值=阴性对照荧光值×3.5。结论试剂盒各项指标均达到临床检测要求,可替代酶免法试剂盒。

Objective To prepare a time-resolved fluoroimmunoassay （TRFIA） kit for clinical detection oflgM antibodies to hepatitis B core antigen （HBc）. Methods Immunocapture method was used to develop the TRFIA kit for detection of the anti-HBc IgM antibodies, and the precision, cross-reactivity and sensitivity of the kit were tested with the clinical serum samples. Results The intra- and inter-assay coefficients of variation of the TRFIA kit were 4.8%-7.2% and 7.5%-8.6%, respectively, and no cross-reactivity with anti-HBs, anti-HBc-IgG or anti-HBe was found. Comparison of the results of the TRFIA kit and enzyme-linked immunosorbent assay （ELISA） demonstrated greater sensitivity of the kit than ELISA in detecting the anti-HBc IgM antibodies in 584 serum samples. According to the detection results in 300 serum samples from healthy donors, the cutoff value of the TRFIA kit was 4.5 times of the fluorescence value of the negative control. Conclusion This TRFIA kit for detecting anti-HBc IgM antibodies meets the demand for clinical application and can replace the ELISA kits.