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目的研制日本血吸虫SjP38的时间分辨免疫荧光分析(Time-resolved fluoroimmunoassay,TRFIA)试剂盒,并评价其效能。方法将抗SjP38的9G7单克隆抗体作为捕获抗体包被96孔板,Eu~(3+)标记1A6单克隆抗体作为检测抗体,研制SjP38 TRFIA试剂盒,并评价试剂盒的准确度、灵敏度、精密度、稳定性以及与病原学检测方法的符合率等指标。结果自制试剂盒准确度好,线性范围为2~1 250 ng/ml,最低检出浓度为0.14 ng/ml;精密度良好,分析内精密度为3.6%~4.6%,分析间精密度为5.1%~6.7%。定性试验表明该试剂盒可在4℃稳定半年,37℃稳定7 d。自制试剂盒与病原学检测方法的阳性符合率为95%,阴性符合率为100%。结论所研制试剂盒的各项性能及检测指标均达到临床检验的要求,但临床推广使用还有待进一步的验证。
Objective To develop the time-resolved fluoroimmunoassay (TRFIA) kit for SjP38 of Schistosoma japonicum and evaluate its efficacy. Methods SjP38 TRFIA kit was developed by using anti-SjP38 monoclonal antibody 9G7 as a capture antibody coated 96-well plate and Eu 3+ labeled 1A6 monoclonal antibody as detection antibody. The accuracy, sensitivity and precision of the kit were also evaluated Degree, stability and coincidence with the etiological detection methods and other indicators. Results The accuracy of home-made kit was good. The linearity range was 2 ~ 1 250 ng / ml and the minimum detectable concentration was 0.14 ng / ml. The precision was good. The intra-assay precision was 3.6% -4.6% % ~ 6.7%. Qualitative experiments show that the kit can be stable at 4 ℃ for six months, 37 ℃ stable 7 d. The positive coincidence rate of homemade kit with etiological detection method was 95% and the negative coincidence rate was 100%. Conclusion The performance of the developed kit and the detection indexes all reach the requirements of clinical examination, but the clinical application needs to be further verified.