PCR和实时荧光PCR法检测华支睾吸虫的比较研究

目的 利用PCR技术建立快速、灵敏、特异的检测华支睾吸虫的方法,并比较PCR和实时荧光PCR检测华支睾吸虫的灵敏性和特异性,探讨其在华支睾吸虫检测中的应用价值.方法 根据华支睾吸虫的特异性基因序列,设计了PCR引物及实时荧光PCR的特异性引物和探针,分别进行灵敏性和特异性实验.结果 设计的引物能够扩增华支睾吸虫DNA,而且探针的特异性很高.实时荧光PCR对华支睾吸虫的检测阈值为3 fg/μl,灵敏度比PCR高两个数量级.结论 PCR和实时荧光PCR检测华支睾吸虫的灵敏性和特异性均很高,操作简便,为华支睾吸虫的检测提供了有效的技术手段和方法.

Comparative study on detection of Clonorchis sinensis by PCR and real-time PCR

Objective To establish a quick, sensitive and specific method for detecting Clonorchis sinensis and evaluate the sensitivity and specificity of PCR and real-time PCR in detection of C. sinensis.Methods According to the specific sequence of C. sinensis genes, the primers of PCR and real-time PCR were designed separately and the fluorogenic probe was designed and synthesized. Results The designed primers could amplify the DNA of C.sinensis and the probes were of high specificity and stability.The detection thres hold by real-time PCR was 3 fg/μl,and the sensitivity of real-time PCR Was higher than PCR. Conclusion PCR and real-time PCR are both simple.rapid methods with hish sensitivity and specificity.Both of them provide an efficient way for detection of C. sinensis.