Development of a real-time PCR assay for the detection of Clonorchis sinensis DNA in gallbladder bile and stone samples from patients with cholecystolithiasis.

Abstract	High prevalence of cholecystolithiasis in parts of East Asia has been postulated to be associated with Clonorchis sinensis infection. This study describes the development of a TaqMan-based real-time PCR assay for the detection of C. sinensis DNA in gallbladder bile and stone samples from patients with cholecystolithiasis. Primers and probe targeting the cytochrome c oxidase subunit 1 gene of mitochondrial DNA proved to be highly specific for C. sinensis and did not amplify other related heterogeneous DNA samples. The detection limit of this assay was 0.1 pg of adult C. sinensis genomic DNA. All of the egg-positive samples determined by microscopy yielded positive results by real-time PCR assay and that genetic testing of gallbladder stones using real-time PCR was considered as the most effective means for assessing C. sinensis infection status. This assay not only contributes to a greater understanding of stone pathogenesis but also benefits patients with cholecystolithiasis by facilitating effective diagnosis, treatment, and relapse prevention.
Authors	Tie Qiao, Pei-Ming Zheng, Rui-Hong Ma, Xiao-Bing Luo, Zhen-Liang Luo
Journal	Parasitology research (Parasitol Res) Vol. 111 Issue 4 Pg. 1497-503 (Oct 2012) ISSN: 1432-1955 [Electronic] Germany
PMID	22669695 (Publication Type: Evaluation Study, Journal Article)
Chemical References	DNA Primers DNA, Helminth Oligonucleotide Probes Electron Transport Complex IV
Topics	Adult Animals Asia Bile (parasitology) Cholecystolithiasis (complications) Clonorchiasis (diagnosis, parasitology) Clonorchis sinensis (genetics, isolation & purification) DNA Primers (genetics) DNA, Helminth (genetics, isolation & purification) Electron Transport Complex IV (genetics) Female Gallbladder (parasitology) Humans Male Middle Aged Molecular Diagnostic Techniques (methods) Oligonucleotide Probes (genetics) Parasitology (methods) Real-Time Polymerase Chain Reaction (methods) Sensitivity and Specificity

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