Journal
KOREAN JOURNAL OF PARASITOLOGY
Volume 51, Issue 6, Pages 645-650Publisher
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
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Funding
- National Science and Technology Development Agency
- Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission through the Health Cluster (SHeP-GMS)
- Khon Kaen University, Thailand
- 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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