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 P
2013; Korean Society for Parasitology; Volume: 51; Issue: 6 Linguagem: Inglês
10.3347/kjp.2013.51.6.645
ISSN1738-0006
AutoresTongjit Thanchomnang, Pewpan M. Intapan, Chairat Tantrawatpan, Viraphong Lulitanond, Sudchit Chungpivat, Piyanan Taweethavonsawat, Worasak Kaewkong, Oranuch Sanpool, Penchom Janwan, Wej Choochote, Wanchai Maleewong,
Tópico(s)Mosquito-borne diseases and control
ResumoA 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℃, 79.0±0.3℃, 76.8±0.1℃, and 79.9±0.1℃, 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. Key words: Wuchereria bancrofti, Brugia malayi, Dirofilaria immitis, Brugia pahangi, high resolution melting real-time PCR, dog, mosquito
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