title: Development of a TaqMan based real-time PCR assay for detection of Clonorchis sinensis DNA in human stool samples and fishes.
abstract: Clonorchiasis caused by the oriental liver fluke Clonorchis sinensis is a fish-borne zoonosis endemic in a number of countries. This article describes the development of a TaqMan based real-time PCR assay for detection of C. sinensis DNA in human feces and in fishes. Primers targeting the first internal transcribed spacer (ITS-1) sequence of the fluke were highly specific for C. sinensis, as evidenced by the negative amplification of closely related trematodes in the test with the exception of Opisthorchis viverrini. The detection limit of the assay was 1pg of purified genomic DNA, 5EPG (eggs per gram feces) or one metacercaria per gram fish filet. The assay was evaluated by testing 22 human fecal samples and 37 fish tissues microscopically determined beforehand, and the PCR results were highly in agreement with the microscopic results. This real-time PCR assay provides a useful tool for the sensitive detection of C. sinensis DNA in human stool and aquatic samples in China and other endemic countries where O. viverrini and Opisthorchis felineus are absent.
authors: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, Gansu Province 730046, China.
authors: Xian-Quan
authors: XQ
authors: Cai
authors: Hai-Qiong
authors: HQ
authors: Yu
authors: Jian-Shan
authors: JS
authors: Bai
authors: Jian-Dong
authors: JD
authors: Tang
authors: Xu-Chu
authors: XC
authors: Hu
authors: Ding-Hu
authors: DH
authors: Chen
authors: Ren-Li
authors: RL
authors: Zhang
authors: Mu-Xin
authors: MX
authors: Chen
authors: Lin
authors: L
authors: Ai
authors: Xing-Quan
authors: XQ
authors: Zhu
mesh_terms: D000818:Animals; D002371:Cat Diseases / Q000469:parasitology; D002415:Cats; D002681:China; D003003:Clonorchiasis / Q000175:diagnosis*; D003004:Clonorchis sinensis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D018509:DNA, Helminth / Q000235:genetics / Q000302:isolation & purification*; D021903:DNA, Ribosomal Spacer; D005243:Feces / Q000469:parasitology*; D005393:Fish Diseases / Q000469:parasitology; D005399:Fishes; D006801:Humans; D010063:Ovum / Q000254:growth & development; D010274:Parasitology / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D018509:DNA, Helminth; D021903:DNA, Ribosomal Spacer