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Development of a PCR-based method for the detection of Opisthorchis viverrini in experimentally infected hamsters

Published online by Cambridge University Press:  21 May 2002

S. WONGRATANACHEEWIN ,
W. PUMIDONMING ,
R.W. SERMSWAN  and
W. MALEEWONG
S. WONGRATANACHEEWIN
Affiliation:
Department of Microbiology, Faculty of Medicine, Khon Kaen University, 123 Mitraparb Road, Khon Kaen, 40002, Thailand
W. PUMIDONMING
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, 123 Mitraparb Road, Khon Kaen, 40002, Thailand
R.W. SERMSWAN
Affiliation:
Department of Biotechnology, Faculty of Technology, Khon Kaen University, 123 Mitraparb Road, Khon Kaen, 40002, Thailand
W. MALEEWONG
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, 123 Mitraparb Road, Khon Kaen, 40002, Thailand
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Abstract

Opisthorchis viverrini infection is an endemic disease that causes a serious public health problem in southeast Asia, especially in northeast Thailand. We have developed a PCR method using a pair of primers named OV-6F/OV-6R for detecting O. viverrini eggs in stool samples and compared it with Stoll's egg-count method. The primers were designed based on the pOV-A6 specific DNA probe sequence which gave a 330 base pair product. The PCR method can detect a single egg in artificially inoculated faeces or as little as 2×10-17ng of O. viverrini genomic DNA. The method gave 100% sensitivity in all hamster groups except in animals exposed to the lowest intensity of infection (1 metacercaria/hamster). In the first month of infection, the PCR method was more sensitive than using the egg-count method in all infected groups especially in the light infections. The PCR method was also successfully used in monitoring a therapeutic study. Since the PCR method showed no cross-reaction with Heterophyid flukes, it can be useful for specific identification of O. viverrini eggs in stool samples without the risk of false positives. It also has great potential for application in clinical epidemiological studies.

Keywords

Opisthorchis viverriniPCRdetection specific DNA probehamster
Type
Research Article
Information
Parasitology , Volume 122 , Issue 2 , February 2001 , pp. 175 - 180
Copyright
© 2001 Cambridge University Press

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