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

• Main Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Chairat Tantrawatpan, Viraphong Lulitanond, Sudchit Chungpivat, Piyanan Taweethavonsawat, Worasak Kaewkong, Oranuch Sanpool, Penchom Janwan, Wej Choochote, Wanchai Maleewong
• Format: Journal
• Published: 2018
• Subjects: Immunology and Microbiology; Medicine
• Online Access: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84893315121&origin=inward; http://cmuir.cmu.ac.th/jspui/handle/6653943832/52622
• Institution: Chiang Mai University

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. © 2013, Korean Society for Parasitology and Tropical Medicine.