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Detection of Opisthorchis viverrini in experimentally infected bithynid snails and cyprinoid fishes by a PCR-based method

Published online by Cambridge University Press:  17 February 2003

W. MALEEWONG ,
P. M. INTAPAN ,
C. WONGKHAM ,
T. WONGSAROJ ,
T. KOWSUWAN ,
W. PUMIDONMING ,
P. PONGSASKULCHOTI  and
V. KITIKOON
W. MALEEWONG
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
P. M. INTAPAN
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
C. WONGKHAM
Affiliation:
Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
T. WONGSAROJ
Affiliation:
Division of General Communicable Diseases, Department of Communicable Disease Control, Ministry of Public Health, Nonthaburi 11000, Thailand
T. KOWSUWAN
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
W. PUMIDONMING
Affiliation:
Department of Microbiology and Parasitology, Faculty of Medical Science, Naresuan University, Pitsanuloke 65000, Thailand
P. PONGSASKULCHOTI
Affiliation:
Applied Malacology Centre, Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
V. KITIKOON
Affiliation:
Applied Malacology Centre, Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
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Abstract

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A PCR procedure for the detection of Opisthorchis viverrini in experimentally infected bithynid snails and cyprinoid fishes was developed. This procedure was based on primers designed from a pOV-A6 specific probe sequence giving a 330 base-pair product. The detection was accomplished in host tissue homogenates to which a single cercaria or metacercaria was introduced. PCR can detect as little as a single cercaria artificially inoculated in a snail or a single metacercaria artificially inoculated in a fish sample. The method gave a 100% positivity rate for all infected snails or fishes. The method did not yield a 330 base-pair amplified product with other digenean fluke DNAs such as Haplorchis taichui, Centrocestus spp., Echinostoma malayanum, Fasciola gigantica, animal schistosomes, Paragonimus heterotremus or Haplorchoides spp. The assay has great potential for application in epidemiological surveys of both snail and fish intermediate hosts as well as for investigation of foodborne parasites in freshwater fishes.

Keywords

Opisthorchis viverriniPCRdetectionbithynid snailcyprinoid fish
Type
Research Article
Information
Parasitology , Volume 126 , Issue 1 , January 2003 , pp. 63 - 67
Copyright
2003 Cambridge University Press