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Development of a real-time PCR assay with fluorophore-labelled hybridization probes for detection of Schistosoma mekongi in infected snails and rat feces

Published online by Cambridge University Press:  01 May 2012

O. SANPOOL ,
P. M. INTAPAN ,
T. THANCHOMNANG ,
P. SRI-AROON ,
V. LULITANOND ,
L. SADAOW  and
W. MALEEWONG
O. SANPOOL
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
P. M. INTAPAN*
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
T. THANCHOMNANG
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Faculty of Medicine, Mahasarakham University, Mahasarakham 44000, Thailand
P. SRI-AROON
Affiliation:
Applied Malacology Center, Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
V. LULITANOND
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
L. SADAOW
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
W. MALEEWONG
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
*
*Corresponding author: Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. Tel: +66 43 348387. Fax: +66 43202475. E-mail: [email protected]
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Summary

Schistosoma mekongi, a blood-dwelling fluke, is a water-borne parasite that is found in communities along the lower Mekong River basin, i.e. Cambodia and Lao People's Democratic Republic. This study developed a real-time PCR assay combined with melting-curve analysis to detect S. mekongi in laboratory setting conditions, in experimentally infected snails, and in fecal samples of infected rats. The procedure is based on melting-curve analysis of a hybrid between an amplicon from S. mekongi mitochondrion sequence, the 260 bp sequence specific to S. mekongi, and specific fluorophore-labelled probes. This method could detect as little as a single cercaria artificially introduced into a pool of 10 non-infected snails, a single cercaria in filtered paper, and 2 eggs inoculated in 100 mg of non-infected rat feces. All S. mekongi-infected snails and fecal samples from infected rats were positive. Non-infected snails, non-infected rat feces, and genomic DNA of other parasites were negative. The method gave high sensitivity and specificity, and could be applied as a fast and reliable tool for cercarial location in water environments in endemic areas and for epidemiological studies and eradication programmes for intermediate hosts.

Keywords

fecesratreal-time PCRSchistosoma mekongisnails
Type
Research Article
Information
Parasitology , Volume 139 , Issue 10 , September 2012 , pp. 1266 - 1272
Copyright
Copyright © Cambridge University Press 2012

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