华支睾吸虫ICT和PCR检测方法的建立及其应用研究

发布时间：2018-03-26 13:02

本文选题：华支睾吸虫　切入点：免疫胶体金　出处：《中国热带医学》2016年12期

【摘要】：目的建立华支睾吸虫的免疫胶体金(ICT)和实时荧光定量PCR检测体系,比较两者检测华支睾吸虫的效果。方法以华支睾吸虫成虫抗原包被胶体金,检测血液样本中的相应抗体,建立华支睾吸虫ICT检测方法;以华支睾吸虫18S r RNA基因作为靶基因,设计RT-PCR的特异性引物和探针,检测粪便样本中的核酸实时荧光定量PCR方法;以两种检测体系与商品化的酶联免疫(ELISA)检测试剂盒平行测定比较,考核检测体系的应用价值。结果对200份华支睾吸虫重点人群样本进行检测,ELISA法检出的华支睾吸虫Ig G抗体阳性例数为20例,阳性率为10.0%;ICT法为16例,阳性率为8.0%,灵敏度为80.0%,特异度为100.0%,符合率为98.0%,ICT和ELISA两种检测方法差异无统计学意义(χ2=2.25,P0.05);RT-PCR法检出的粪便样本中华支睾吸虫核酸阳性例数为18例,阳性率为9.0%,灵敏度为90.0%,特异度为100.0%,符合率为99.0%,RT-PCR和ELISA两种检测方法差异无统计学意义(χ2=0.05,P0.05)。说明ICT、RT-PCR两种检测方法与ELISA的测定结果比较一致。结论建立的ICT及RT-PCR两种检测方法特异性好、灵敏度高,两种检测方法均适用于华支睾吸虫现场的快速检测及定量分析。
[Abstract]:Objective to establish an immunoassay system for clonorchiasis sinensis (ICTT) and real-time fluorescent quantitative PCR (PCR) to compare the effect of the two methods in detecting clonorchiasis sinensis. Methods Colloidal gold was coated with adult worm antigen of Clonorchis sinensis to detect the corresponding antibodies in blood samples. A ICT detection method for Clonorchis sinensis was established, and the 18s r RNA gene of Clonorchis sinensis was used as the target gene. The specific primers and probes of RT-PCR were designed to detect the real-time fluorescence quantitative PCR method of nucleic acid in fecal samples. Two detection systems were compared with commercial Elisa Elisa kit. Results among 200 samples of clonorchiasis sinensis detected by Elisa, 20 cases were positive for IgG antibody, and the positive rate was 10.0% ICT method in 16 cases. The positive rate was 8.0%, the sensitivity was 80.0 and the specificity was 100.0.The coincidence rate was 98.0%. There was no significant difference between the two methods (蠂 2 / 2.25P 0.05 / RT-PCR). 18 cases were positive for nucleic acid of Clonorchis sinensis. The positive rate was 9.0%, the sensitivity was 90.0 and the specificity was 100.0.The coincidence rate was 99.0%. There was no significant difference between RT-PCR and ELISA (蠂 ~ 2 / 0.05) (蠂 ~ 2 / 0.05). The results showed that the results of RT-PCR and RT-PCR were consistent with those of ELISA. Conclusion the established ICT and RT-PCR methods have good specificity. The two methods are suitable for rapid detection and quantitative analysis of Clonorchis sinensis.
【作者单位】：深圳市疾病预防控制中心;南华大学公共卫生学院;
【基金】：国家卫生行业科研专项(No.201502021)
【分类号】：R532.23;R440

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