title: A specific PCR assay for the diagnosis of Clonorchis sinensis infection in humans, cats and fishes.
abstract: Clonorchiasis caused by Clonorchis sinensis is a fish-borne parasitic disease which is endemic in a number of countries. Using the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of C. sinensis as genetic markers, a pair of C. sinensis-specific primers was designed and used to establish a specific PCR assay for the diagnosis of C. sinensis infection in humans, cats and fish. This approach allowed the specific identification of C. sinensis after optimizing amplification conditions, with no amplicons being amplified from related heterogeneous DNA samples, and sequencing of amplicons confirmed the identity of the sequences amplified. The detection limit of this assay was 1.03 pg of adult C. sinensis, 1.1 metacercariae per gram of fish filet, and a single egg in human and cat feces. The PCR assay should provide a useful tool for the diagnosis and molecular epidemiological investigation of clonorchiasis in humans and animals.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, Gansu Province 730046, PR China.
authors.forename: Si-Yang
authors.initials: SY
authors.lastname: Huang
authors.forename: Jian-Dong
authors.initials: JD
authors.lastname: Tang
authors.forename: Hui-Qun
authors.initials: HQ
authors.lastname: Song
authors.forename: Bao-Quan
authors.initials: BQ
authors.lastname: Fu
authors.forename: Min-Jun
authors.initials: MJ
authors.lastname: Xu
authors.forename: Xu-Chu
authors.initials: XC
authors.lastname: Hu
authors.forename: Han
authors.initials: H
authors.lastname: Zhang
authors.forename: Ya-Biao
authors.initials: YB
authors.lastname: Weng
authors.forename: Rui-Qing
authors.initials: RQ
authors.lastname: Lin
authors.forename: Xing-Quan
authors.initials: XQ
authors.lastname: Zhu
mesh_terms: D000818:Animals; D002371:Cat Diseases / Q000469:parasitology*; D002415:Cats; 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; D058488:Metacercariae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D010063:Ovum / Q000254:growth & development; D010274:Parasitology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D018509:DNA, Helminth; D021903:DNA, Ribosomal Spacer