Pusadee Sri-AroonPewpan M. IntapanChantima LohachitPhunthira PhongsasakulchotiTongjit ThanchomnangViraphong LulitanondAlexandra HiscoxSamlane PhompidaPany SananikhomWanchai MaleewongPaul T. BreyMahidol UniversityKhon Kaen UniversityMahasarakham UniversityInstitut Pasteur du LaosMinistry of HealthNam Theun 2 Power Company2018-05-032018-05-032011-04-01Parasitology Research. Vol.108, No.4 (2011), 973-97814321955093201132-s2.0-79956207694https://repository.li.mahidol.ac.th/handle/20.500.14594/11339Naturally 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.Mahidol UniversityAgricultural and Biological SciencesImmunology and MicrobiologyMedicineVeterinaryArticleSCOPUS10.1007/s00436-010-2140-3