Real-time fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) supplemented
with melting curve analysis is a highly sensitive and fast method offering a high throughput. We report the development
of a real-time FRET PCR for molecular detection of Opisthorchis viverrini in human fecal samples. The diagnostic sensitivity,
specificity, accuracy, and positive and negative predictive values of this method were 97.5%, 100%, 98.9%, 100%,
and 98.2%, respectively. The sensitivity was not significantly different from that of the quantified formalin-ethyl acetate
concentration technique, the gold standard ( P > 0.05). The procedure has potential for diagnosis of human opisthorchiasis
in disease-endemic areas, for large epidemiologic investigations involving at risk populations, and monitoring eradication
programs of the liver fluke, which causes hepatobiliary diseases and induces cholangiocarcinoma.