Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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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)
  • Received : 2013.05.30
  • Accepted : 2013.10.11
  • Published : 2013.12.31
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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

References

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