Chin J Schisto Control ›› 2017, Vol. 29 ›› Issue (1): 64-67.

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Development and effectiveness evaluation of time-resolved fluoroimmunoassay kit for detection of Schistosoma japonicum

MENG Qing-dong1| WANG Yan2| SUN Wen-qiao3| REN Shou-lei3| XIN Bo3| ZHU Peng-fei3| LI Kang-yan4| LIANG Huan-kun4| ZHANG Li-cheng3*   

  1. 1 Shandong Provincial Institute of Parasitic Diseases| Third Affiliated Hospital of Shandong Province Academy of Medical Sciences| Jining 272033| China; 2 Shandong Medicine Technician College| China; 3 The 88th Hospital of PLA| China; 4 Guangzhou Youdi Biological Technology Co.| Ltd.| China
  • Online:2017-02-23 Published:2017-02-24
  • Contact: ZHANG Li?cheng

血吸虫病时间分辨免疫荧光检测试剂盒的研制和效能评价

孟庆东1|王燕2|孙文俏3|任守雷3|信波3|朱鹏飞3|李康燕4|梁焕坤4|张立成3*   

  1. 1 山东省寄生虫病防治研究所|山东省医学科学院第三附属医院(济宁272033); 2 山东医药技师学院; 3中国人民解放军第88医院; 4 广州优迪生物科技有限公司
  • 通讯作者: 张立成
  • 作者简介:孟庆东|男|硕士研究生|主治医生。研究方向:胃肠外科
  • 基金资助:
    广东省省级科技型中小企业技术创新专项资金项目(2006A010119067)

Abstract: Objective To develop a kit of time?resolved fluoroimmunoassay (TRFIA) for detection of Schistosoma japonicum protein SjP38, and evaluate its effectiveness. Methods The anti 9G7 SjP38 monoclonal antibody was used as the capture antibody coated with 96?hole plate, and the Eu3+ labeled 1A6 monoclonal antibody was used as the detection antibody to establish the TRFIA SjP38 kit. In addition, the accuracy, sensitivity, precision, stability and coincidence rate to pathogenic diagnosis of the kit were evaluated. Results This established kit possessed high accuracy, wide linear range from 2 to 1 250 ng/ml, high sensitivity with the minimum detectable concentration of 0.14 ng/ml, and good precision (the coefficient variation of the intra?and inter?assay were 3.6% to 4.6% and 5.1% to 6.7%, respectively). The stability tests showed that the reagents could be stable for six months at 4 ℃, 7 d at 37 ℃. The positive and negative corresponding rates to the pathogen detection method were 95% and 100% respectively. Conclusion All the performance and detection indicators of the kit have reached the requirements of clinical test, but its clinical application still needs further validation.

Key words: Schistosoma japonicum; Recombinant protein SjP38; Double?antibody sandwich; Time?resolved fluoroimmunoassay (TRFIA)

摘要: 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,用Eu3+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。

关键词: 日本血吸虫; 重组蛋白SjP38; 双抗体夹心; 时间分辨免疫荧光分析

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