Molecular evidence of Opisthorchis viverrini in infected bithyniid snails in the Lao People's Democratic Republic by specific hybridization probe-based real-time fluorescence resonance energy transfer PCR method.

Pusadee Sri-Aroon 1 Pewpan M Intapan 2 Chantima Lohachit 1 Phunthira Phongsasakulchoti 1 Tongjit Thanchomnang 3 Viraphong Lulitanond 4 Alexandra Hiscox 5 Samlane Phompida 6 Pany Sananikhom 7 Wanchai Maleewong 2, 8, * Paul T Brey 5
* Corresponding author
1 Applied Malacology Center
2 Department of Parasitology
3 Faculty of Medicine
4 Department of Microbiology
5 Institut Pasteur du Laos
6 National Center for Malariology, Parasitology and Entomology
7 Health Program Management Unit
8 Department of Parasitology
Abstract : Naturally occurring bithyniid snails, Bithynia siamensis goniomphalos (Prosobranchia: Bithyniidae), and their intermediate hosts were sampled from Khammouane Province, the Lao People's Democratic Republic, and the prevalence of the carcinogenic human liver fluke, Opisthorchis viverrini, was examined. The presence of O. viverrini cercariae in snails was examined by cercarial shedding test and then confirmed by specific hybridization probe-based real-time fluorescence resonance energy transfer (FRET) PCR method. The real-time FRET PCR method is based on a fluorescence melting curve analysis of a hybrid between an amplicon produced from the pOV-A6 specific sequence (Genbank accession no. S80278), a 162-bp repeated sequence specific to O. viverrini, and specific fluorophore-labeled probes. Mean melting temperature of O. viverrini DNA from the cercariae and each of two positive snails by shedding test was 66.3 ± 0.1. The O. viverrini infection rate in snails was 2.47% (2/81) by cercarial shedding test but was 8.52% (4/47) by real-time FRET PCR method. The real-time FRET PCR method is rapid and effective in examining a large number of snail samples simultaneously. Validation using molecular evidence from this procedure provides another tool for surveying the prevalence of O. viverrini-infected snails in Southeast Asian countries.
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Submitted on : Thursday, June 6, 2013 - 10:57:27 AM
Last modification on : Thursday, March 7, 2019 - 3:04:03 PM

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Pusadee Sri-Aroon, Pewpan M Intapan, Chantima Lohachit, Phunthira Phongsasakulchoti, Tongjit Thanchomnang, et al.. Molecular evidence of Opisthorchis viverrini in infected bithyniid snails in the Lao People's Democratic Republic by specific hybridization probe-based real-time fluorescence resonance energy transfer PCR method.. Parasitology Research, Springer Verlag (Germany), 2011, 108 (4), pp.973-8. ⟨10.1007/s00436-010-2140-3⟩. ⟨pasteur-00830974⟩

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