title: Molecular survey of Dirofilaria immitis and Dirofilaria repens by new real-time TaqMan®PCR assay in dogs and mosquitoes (Diptera: Culicidae) in Corsica (France).
abstract: Dirofilaria immitis and D. repens are filarioid nematodes of animals and humans, transmitted by the bite of infected mosquitoes. Domestic and wild canids are a major natural host and reservoir for these parasites. In this study, we designed a duplex real-time PCR protocol targeting the mitochondrial cytochrome c oxidase subunit I (COI) gene, detecting both D. immitis and D. repens using two primer pairs and two Dirofilaria-specific hydrolysable probes. The sensitivity and specificity of the primers and probes were tested in both experimental and naturally infected samples. The detection limits of this assay were evaluated using plasmid DNA from D. immitis and D. repens. No cross-reaction was observed when testing this system against DNA from other filarial nematodes. The detection limit of the real-time PCR system was one copy per reaction mixture containing 5μl of template DNA. Field application of the new duplex real-time assay was conducted in Corsica. The prevalence rate of D. immitis was 21.3% (20/94) in dogs. In a locality where most dogs with Dirofilaria spp. infection were found, D. immitis and D. repens were detected in 5% (20/389) and 1.5% (6/389) of the Aedes albopictus population, respectively. These results suggest that this sensitive assay is a powerful tool for monitoring dirofilariosis in endemic or high risk areas.
authors.affiliation: Research Unit of Emerging Infectious and Tropical Diseases (URMITE) UMR CNRS 7278 IRD 198 INSERM U1015 Aix-Marseille University, Marseille, France.
authors.forename: Djamel
authors.initials: D
authors.lastname: Tahir
authors.affiliation: Research Unit of Emerging Infectious and Tropical Diseases (URMITE) UMR CNRS 7278 IRD 198 INSERM U1015 Aix-Marseille University, Marseille, France.
authors.forename: Fadi
authors.initials: F
authors.lastname: Bittar
authors.affiliation: OCIC, Office de l'Environnement de la Corse, Corte, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Barré-Cardi
authors.affiliation: Research Unit of Emerging Infectious and Tropical Diseases (URMITE) UMR CNRS 7278 IRD 198 INSERM U1015 Aix-Marseille University, Marseille, France.
authors.forename: Doudou
authors.initials: D
authors.lastname: Sow
authors.affiliation: Research Unit of Emerging Infectious and Tropical Diseases (URMITE) UMR CNRS 7278 IRD 198 INSERM U1015 Aix-Marseille University, Marseille, France.
authors.forename: Mustapha
authors.initials: M
authors.lastname: Dahmani
authors.affiliation: Research Unit of Emerging Infectious and Tropical Diseases (URMITE) UMR CNRS 7278 IRD 198 INSERM U1015 Aix-Marseille University, Marseille, France.
authors.forename: Oleg
authors.initials: O
authors.lastname: Mediannikov
authors.affiliation: Research Unit of Emerging Infectious and Tropical Diseases (URMITE) UMR CNRS 7278 IRD 198 INSERM U1015 Aix-Marseille University, Marseille, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Raoult
authors.affiliation: Research Unit of Emerging Infectious and Tropical Diseases (URMITE) UMR CNRS 7278 IRD 198 INSERM U1015 Aix-Marseille University, Marseille, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Davoust
authors.affiliation: Research Unit of Emerging Infectious and Tropical Diseases (URMITE) UMR CNRS 7278 IRD 198 INSERM U1015 Aix-Marseille University, Marseille, France. Electronic address: philippe.parola@univ-amu.fr.
authors.forename: Philippe
authors.initials: P
authors.lastname: Parola
mesh_terms: D000330:Aedes / Q000469:parasitology; D000818:Animals; D009033:Culicidae / Q000469:parasitology*; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D004183:Dirofilaria immitis / Q000235:genetics / Q000302:isolation & purification*; D060348:Dirofilaria repens / Q000235:genetics / Q000302:isolation & purification*; D004184:Dirofilariasis / Q000453:epidemiology / Q000469:parasitology*; D004283:Dog Diseases / Q000453:epidemiology / Q000469:parasitology*; D004285:Dogs; D005260:Female; D005602:France / Q000453:epidemiology; D007303:Insect Vectors / Q000469:parasitology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D018509:DNA, Helminth