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

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
Sri-Aroon, P; Intapan, PM; Lohachit, C; Phongsasakulchoti, P; Thanchomnang, T; Lulitanond, V; Hiscox, A; Phompida, S; Sananikhom, P; Maleewong, W; Brey, PT
Keywords
MELTING CURVE ANALYSIS; CLONORCHIS-SINENSIS; LIVER FLUKE; PARASITE INFECTIONS; THAILAND; CHOLANGIOCARCINOMA; PDR; PRAZIQUANTEL; FISHES
Issue Date
2011
Publisher
PARASITOL RES
Citation
Parasitol. Res.;APR;2011;108;4
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.
URI
http://hdl.handle.net/11267/2546
ISSN
0932-0113
Appears in Collections:
2.Ministry of Health > Journal articles
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