华支睾吸虫病患者粪便虫卵核糖体DNA内转录间隔区序列分析

Ribosomal DNA internal transcribed spacer sequencing for identification of Clonorchis sinensis eggs in stool from human cases

目的 通过PCR方法扩增华支睾吸虫病患者粪便虫卵核糖体DNA （ribosomal DNA,rDNA）内转录间隔区（internal transcribed spacer,ITS）序列进行分析,探讨其用于华支睾吸虫感染检测的价值. 方法 提取2例华支睾吸虫患者粪便中的虫卵DNA作模板,以rDNA ITS基因片段为目的片段进行PCR扩增,对扩增片段进行测序分析. 结果 从2例患者粪便虫卵样本中均扩增出1 123 bp的条带,经序列比对确定患者粪便中的虫卵为华支睾吸虫卵.测序分析发现本研究的两个样本与中国黑龙江分离株（KF740425）相似性为100％. 结论 本研究从2例华支睾吸虫病患者粪便虫卵中成功扩增出rDNA ITS基因,为从分子生物学角度检测华支睾吸虫病提供了资料.

Objective To amplify and analyse the internal transcribed spacer（ITS） of ribosomal DNA of Clonorchis sinensis eggs from patients by PCR method so as to identify the infection and explore the detective value of the method.Methods ITS sequence was chosen as target fragment and PCR was used to amplify the DNA of eggs from two patients,and then sequencing was done.Results The length of the ITS sequences was 1 123 bp from the egg samples of two patients.The eggs was confirmed as C.sinensis.The two isolated samples showed 100％ similarity with the specimen from Heilongjiang （KF740425）.Conclusion In this study,the rDNA ITS of egg samples from two clonorchiasis cases was identified by PCR successfully thus to provide information for detecting human clonorchiasis.