论文部分内容阅读
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,P>0.05);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,P>0.05)。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。
Objective To establish the immune colloidal gold (ICT) and real-time fluorescence quantitative PCR (PCR) detection system of Clonorchis sinensis, and compare the detection results of Clonorchis sinensis with both. Methods Clonorchis sinensis adult worm antigen coated with colloidal gold, blood samples were tested for the corresponding antibodies to establish the ICT test method of Clonorchis sinensis; Clonorchis sinensis 18S r RNA gene as a target gene designed RT-PCR specificity Primers and probes to detect stool samples by real-time fluorescence quantitative PCR method; the two detection systems and commercial ELISA kit for parallel determination compared to assess the value of the detection system. Results A total of 200 samples of clonorchiasis key population were detected. The number of IgA positive cases of Clonorchis sinensis detected by ELISA was 20 cases, the positive rate was 10.0%. ICT method was 16 cases, the positive rate was 8.0% , The sensitivity was 80.0%, the specificity was 100.0%, the coincidence rate was 98.0%, there was no significant difference between ICT and ELISA (χ2 = 2.25, P> 0.05); stool samples detected by RT- Clonorchis sinensis nucleic acid positive cases were 18 cases, the positive rate was 9.0%, the sensitivity was 90.0%, the specificity was 100.0%, the coincidence rate was 99.0%, RT-PCR and ELISA two detection methods was no significant difference (χ2 = 0.05, P> 0.05). Indicating ICT and RT-PCR two detection methods and ELISA determination of the results are more consistent. Conclusion The established detection methods of ICT and RT-PCR have good specificity and high sensitivity. Both detection methods are suitable for the rapid detection and quantitative analysis of Clonorchis sinensis.