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.
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℃, 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.
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Faculty of Medicine, Mahasarakham University, Mahasarakham 44000, Thailand.
authors: Tongjit
authors: T
authors: Thanchomnang
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Pewpan M
authors: PM
authors: Intapan
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Division of Cell Biology, Department of Preclinical Sciences, Faculty of Medicine, Thammasat University, Rangsit Campus, Pathum Thani 12121, Thailand.
authors: Chairat
authors: C
authors: Tantrawatpan
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Viraphong
authors: V
authors: Lulitanond
authors: Parasitology Unit, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.
authors: Sudchit
authors: S
authors: Chungpivat
authors: Parasitology Unit, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.
authors: Piyanan
authors: P
authors: Taweethavonsawat
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Department of Biochemistry, Faculty of Medical Science, Naresuan University, Phitsanulok 65000, Thailand.
authors: Worasak
authors: W
authors: Kaewkong
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Oranuch
authors: O
authors: Sanpool
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Penchom
authors: P
authors: Janwan
authors: Department of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
authors: Wej
authors: W
authors: Choochote
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Wanchai
authors: W
authors: Maleewong
mesh_terms: D000818:Animals; D001769:Blood / Q000469:parasitology*; D002009:Brugia / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002415:Cats; D009033:Culicidae / Q000469:parasitology*; D004183:Dirofilaria immitis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004285:Dogs; D006801:Humans; D008297:Male; D010274:Parasitology / Q000379:methods*; D018510:RNA, Helminth / Q000235:genetics; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D044366:Transition Temperature; D014958:Wuchereria bancrofti / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D018510:RNA, Helminth; D012341:RNA, Ribosomal, 5S
keywords: Brugia malayi; Brugia pahangi; Dirofilaria immitis; Wuchereria bancrofti; dog; high resolution melting real-time PCR; mosquito