A specific PCR assay for the diagnosis of Clonorchis sinensis infection in humans, cats and fishes |
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| Authors: | Huang Si-Yang Tang Jian-Dong Song Hui-Qun Fu Bao-Quan Xu Min-Jun Hu Xu-Chu Zhang Han Weng Ya-Biao Lin Rui-Qing Zhu Xing-Quan |
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| Institution: | a 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 Chinab College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, PR Chinac Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong Province 510642, PR Chinad Guangdong Institute for Animal Disease Control and Surveillance, Guangzhou, Guangdong Province 510230, PR Chinae College of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan Province 650201, PR Chinaf College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang Province 163319, PR China |
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| 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. |
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| Keywords: | Clonorchis sinensis Clonorchiasis Internal transcribed spacers (ITS) PCR assay Detection Human Cat Fish |
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