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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.
Received May 23, 2009. Accepted for publication July 15, 2009.
Acknowledgment: We thank Dr. Mark Roselieb for his assistance with the manuscript preparation.
Financial support: This study was supported by the Thailand Research Fund (grant no. BRG50800005).
* Address correspondence to Pewpan M. Intapan, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. E-mail: pewpan{at}kku.ac.th
Authors addresses: Pewpan M. Intapan, Tongjit Thanchomnang, Viraphong Lulitanond, and Wanchai Maleewong, Departments of Parasitology and Microbiology, Faculty of Medicine and Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand. Phunthira Pongsaskulchoti, Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.
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