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- Title
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.
- Authors
Tongjit Thanchomnang; Pewpan M. Intapan; Chairat Tantrawatpan; Viraphong Lulitanond; Sudchit Chungpivat; Piyanan Taweethavonsawat; Worasak Kaewkong; Oranuch Sanpool; Penchom Janwan; Wej Choochote; Wanchai Maleewong
- 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±0.2°C, 79.0±0.3°C, 76.8 ± 0.1 °C, and 79.9 ± 0.1 °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.
- Subjects
BRUGIA malayi; DIROFILARIA immitis; MOSQUITO vectors; BLOOD sampling; POLYMERASE chain reaction; NEMATODE identification; RIBOSOMAL RNA
- Publication
Korean Journal of Parasitology, 2013, Vol 51, Issue 6, p645
- ISSN
0023-4001
- Publication type
Article
- DOI
10.3347/kjp.2013.51.6.645