3.9 Article

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

Journal

KOREAN JOURNAL OF PARASITOLOGY
Volume 51, Issue 6, Pages 645-650

Publisher

KOREAN SOC PARASITOLOGY, SEOUL NATL UNIV COLL MEDI
DOI: 10.3347/kjp.2013.51.6.645

Keywords

Wuchereria bancrofti; Brugia malayi; Dirofilaria immitis; Brugia pahangi; high resolution melting real-time PCR; dog; mosquito

Categories

Funding

  1. National Science and Technology Development Agency
  2. Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission through the Health Cluster (SHeP-GMS)
  3. Khon Kaen University, Thailand
  4. TRF Senior Research Scholar Grant, Thailand Research Fund [RTA5580004]

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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 +/- 0.2 degrees C, 79.0 +/- 0.3 degrees C, 76.8 +/- 0.1 degrees C, and 79.9 +/- 0.1 degrees 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.

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