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Title: 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
Authors: Pusadee Sri-Aroon
Pewpan M. Intapan
Chantima Lohachit
Phunthira Phongsasakulchoti
Tongjit Thanchomnang
Viraphong Lulitanond
Alexandra Hiscox
Samlane Phompida
Pany Sananikhom
Wanchai Maleewong
Paul T. Brey
Mahidol University
Khon Kaen University
Mahasarakham University
Institut Pasteur du Laos
Ministry of Health
Nam Theun 2 Power Company
Keywords: Agricultural and Biological Sciences;Immunology and Microbiology;Medicine;Veterinary
Issue Date: 1-Apr-2011
Citation: Parasitology Research. Vol.108, No.4 (2011), 973-978
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. © Springer-Verlag 2010.
ISSN: 14321955
Appears in Collections:Scopus 2011-2015

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