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Development and evaluation of loop-mediated isothermal amplification (LAMP) for rapid detection of Clonorchis sinensis from its first intermediate hosts, freshwater snails

Published online by Cambridge University Press:  22 July 2013

Y. CHEN ,
T. WEN ,
D.-H. LAI ,
Y.-Z. WEN ,
Z.-D. WU ,
T.-B. YANG ,
X.-B. YU ,
G. HIDE  and
Z.-R. LUN
Y. CHEN
Affiliation:
Centre for Parasitic Organisms, State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering (Sun Yat-Sen University), Ministry of Education, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, P.R. China
T. WEN
Affiliation:
Centre for Parasitic Organisms, State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering (Sun Yat-Sen University), Ministry of Education, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, P.R. China
D.-H. LAI
Affiliation:
Centre for Parasitic Organisms, State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering (Sun Yat-Sen University), Ministry of Education, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, P.R. China
Y.-Z. WEN
Affiliation:
Centre for Parasitic Organisms, State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering (Sun Yat-Sen University), Ministry of Education, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, P.R. China
Z.-D. WU
Affiliation:
Key Laboratory of Tropical Disease and Control (Sun Yat-Sen University), Ministry of Education, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510080, P.R. China
T.-B. YANG
Affiliation:
Centre for Parasitic Organisms, State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering (Sun Yat-Sen University), Ministry of Education, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, P.R. China
X.-B. YU
Affiliation:
Key Laboratory of Tropical Disease and Control (Sun Yat-Sen University), Ministry of Education, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510080, P.R. China
G. HIDE
Affiliation:
Ecosystems and Environment Research Centre and Biomedical Research Centre, School of Environment and Life Sciences, University of Salford, Salford, M5 4WT, UK
Z.-R. LUN*
Affiliation:
Centre for Parasitic Organisms, State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering (Sun Yat-Sen University), Ministry of Education, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, P.R. China Key Laboratory of Tropical Disease and Control (Sun Yat-Sen University), Ministry of Education, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510080, P.R. China Ecosystems and Environment Research Centre and Biomedical Research Centre, School of Environment and Life Sciences, University of Salford, Salford, M5 4WT, UK
*
*Corresponding author: School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, P.R. China. E-mail: [email protected]
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Summary

Clonorchiasis, caused by Clonorchis sinensis, is a key foodborne zoonosis, which is mainly found in China, Korea and Vietnam. Detection of this parasite from the second intermediate host, the freshwater fish is the common method for epidemiological surveys of this parasite, but is time consuming, labour intensive and easily leads to misdiagnosis. In this study, we have developed a rapid, sensitive and reliable molecular method for the diagnosis of C. sinensis from its first intermediate hosts, freshwater snails, based on a loop-mediated isothermal amplification (LAMP) method. The specific amplified fragment from genomic DNA of C. sinensis did not cross-react with those from other relevant trematodes and a range of hosts (freshwater fish, shrimps and snails) of C. sinensis living in similar environments. The detection limit of the LAMP method was as low as 10 fg which was 1000 times more sensitive than conventional PCR, which was also demonstrated by successful application to field samples. These results show that the LAMP method is a more sensitive tool than conventional PCR for the detection of C. sinensis infection in the first intermediate hosts and, due to a simpler protocol, is an ideal molecular method for field-based epidemiological surveys of this parasite.

Keywords

Clonorchis sinensisloop mediated isothermal amplification (LAMP)first intermediate hostITS2field samplesrapid detection
Type
Research Article
Information
Parasitology , Volume 140 , Issue 11 , September 2013 , pp. 1377 - 1383
Copyright
Copyright © Cambridge University Press 2013 

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