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Article de Anglais | WPRIM | ID: wpr-118761

RÉSUMÉ

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+/-0.2degrees C, 79.0+/-0.3degrees C, 76.8+/-0.1degrees C, and 79.9+/-0.1degrees 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.


Sujet(s)
Animaux , Chats , Chiens , Humains , Mâle , Sang/parasitologie , Brugia/classification , Culicidae/parasitologie , Dirofilaria immitis/classification , Parasitologie/méthodes , ARN des helminthes/génétique , ARN ribosomique 5S/génétique , Réaction de polymérisation en chaine en temps réel/méthodes , Sensibilité et spécificité , Température de transition , Wuchereria bancrofti/classification
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