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http://dx.doi.org/10.3347/kjp.2013.51.6.645

Rapid Detection and Identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in Mosquito Vectors and Blood Samples by High Resolution Melting Real-Time PCR  

Thanchomnang, Tongjit (Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University)
Intapan, Pewpan M. (Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University)
Tantrawatpan, Chairat (Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University)
Lulitanond, Viraphong (Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University)
Chungpivat, Sudchit (Parasitology Unit, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University)
Taweethavonsawat, Piyanan (Parasitology Unit, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University)
Kaewkong, Worasak (Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University)
Sanpool, Oranuch (Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University)
Janwan, Penchom (Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University)
Choochote, Wej (Department of Parasitology, Faculty of Medicine, Chiang Mai University)
Maleewong, Wanchai (Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University)
Publication Information
Parasites, Hosts and Diseases / v.51, no.6, 2013 , pp. 645-650 More about this Journal
Abstract
A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at $81.5{\pm}0.2^{\circ}C$, $79.0{\pm}0.3^{\circ}C$, $76.8{\pm}0.1^{\circ}C$, and $79.9{\pm}0.1^{\circ}C$, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.
Keywords
Wuchereria bancrofti; Brugia malayi; Dirofilaria immitis; Brugia pahangi; high resolution melting real-time PCR; dog; mosquito;
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