乙肝病毒前S1抗原时间分辨荧光免疫分析试剂盒的研制

Development of A Time-Resolved Fluoroimmunoassay Kit for Detection of Hepatitis B Virus preS1 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前S1抗原的检测试剂盒。方法应用双抗体夹心法建立乙肝preS1抗原TRFIA检测试剂盒,对试剂盒的各项性能指标进行评估。结果对280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特异性更好;200份正常人血清样本检测结果表明,该试剂盒的cutoff值=阴性对照荧光值×4.5。用自制质控品检测分析内和分析间的精密度分别为3.4%～7.8%,6.9%～8.4%;检测HBsAg及HBeAg无交叉反应。结论试剂盒各项指标均达到临床检测要求,TRFIA法精密度和特异性有较大提高,可替代酶免法试剂盒。

Objective To evaluate a home-made time-resolved fluoroimmunoassay （TRFIA） kit for the clinical detection of hepatitis B virus preS1 antigen. Methods An antibody-sandwich based TRFIA kit for developed for the detection of hepatitis B virus preS1 antigen. Results 280 serum samples were tested for hepatitis B virus preS1 antigen with the home-made TRFIA kit and the results were compared with the ELISA method. The home-made TRFIA kit was more specific than the ELISA method. According to the results from 200 normal samples, the cutoff value of the home-made kit was 4.5 times of the negative control fluorescence value The intra- and inter-assay coefficient of variation were 3.4%-7.8% and 6.9%-8.4%, respectively. There was no cross reactivity with HBsAg and HBeAg. Conclusion The developed HBV preS1 TRFIA kit is sensitivity and specific for clinical detection of preS1 antigen.