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Development of a real-time PCR assay for the detection of Clonorchis sinensis DNA in gallbladder bile and stone samples from patients with cholecystolithiasis.
- Source :
-
Parasitology research [Parasitol Res] 2012 Oct; Vol. 111 (4), pp. 1497-503. Date of Electronic Publication: 2012 Jun 06. - Publication Year :
- 2012
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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.
- Subjects :
- Adult
Animals
Asia
Clonorchiasis parasitology
Clonorchis sinensis genetics
DNA Primers genetics
DNA, Helminth genetics
Electron Transport Complex IV genetics
Female
Humans
Male
Middle Aged
Molecular Diagnostic Techniques methods
Oligonucleotide Probes genetics
Parasitology methods
Sensitivity and Specificity
Bile parasitology
Cholecystolithiasis complications
Clonorchiasis diagnosis
Clonorchis sinensis isolation & purification
DNA, Helminth isolation & purification
Gallbladder parasitology
Real-Time Polymerase Chain Reaction methods
Subjects
Details
- Language :
- English
- ISSN :
- 1432-1955
- Volume :
- 111
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Parasitology research
- Publication Type :
- Academic Journal
- Accession number :
- 22669695
- Full Text :
- https://doi.org/10.1007/s00436-012-2986-7