Cited by 47Parasitic Diseases Research and TreatmentHelminth infection and controlResearch on Leishmaniasis Studies

A duplex real-time polymerase chain reaction assay for the detection of and differentiation between Dirofilaria immitis and Dirofilaria repens in dogs and mosquitoes

Latrofa, Maria Stefania, Dantas-Torres, Filipe, Annoscia, Giada, Genchi, Marco, Traversa, Donato, Otranto, Domenico·2012·Veterinary Parasitology

Abstract

This study describes a duplex real-time polymerase chain reaction (PCR) assay for the detection and differentiation between Dirofilaria immitis and Dirofilaria repens in dog blood and mosquitoes. Regions of a cytochrome oxidase 1 (cox1) mitochondrial DNA fragment and the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA were amplified from microfilariae and adult worm samples, using a sensitive SsoFast™ EvaGreen(®) based real-time PCR method coupled with melting-curve analysis. The limit of the real-time PCR in detecting microfilaria and adult worm DNA was also tested both in dog blood and in artificially infected microfilarial. Two peaks at different melting temperatures (T(m)) for D. immitis (mean ± SD=75.7 ± 0.3°C) and D. repens (mean ± SD=70 ± 0.7°C), respectively, were obtained for microfilarial and adult positive controls of both species when examined separately and together. The real-time PCR protocol was also efficient in detecting microfilarial and adult DNA of both species when tested in samples spiked with DNA from Aedes albopictus, in Aedes aegypti experimentally infected by D. repens and in Culex pipiens naturally infected by D. repens and D. immitis. The high sensitivity of real-time PCR confirmed its reliability in detecting small amounts of genomic DNA either in dog blood or mosquitoes (2.5 pg/μl and 3 × 10(-1)pg/μl for D. immitis and D. repens, respectively). This assay is proposed as a tool for the epidemiological surveillance of the two most important Dirofilaria species in areas where they are endemic and sympatric.

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Crossref · 10.1016/j.vetpar.2011.10.038

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  • A duplex real-time PCR has been assessed for discrimination between D. immitis and D. repens and their quantification in blood samples and mosquitoes.

    “Recently, a duplex real-time PCR has been assessed for the discrimination between D. immitis and D. repens and their quantification in blood samples and mosquitoes [130].”

    From Vector-borne helminths of dogs and humans in Europe (2013) · cites this paper as [130]

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Authors
Latrofa, M. S., Dantas-Torres, F., Annoscia, G., Genchi, M., Traversa, D., & Otranto, D.
Affiliations
  • University of Bari Aldo Moro (IT)
  • University of Milan (IT)
  • University of Teramo (IT)
Corresponding author
Otranto, Domenico
Venue
Veterinary Parasitology
Publisher
Elsevier BV
Published
2012-04
Volume
185
Issue
2-4
Pages
181-185
ISSN
0304-4017 (ISSN-L)
OpenAlex
W2051126021
Cited by
47
References
41
Citations by year
2026: 1 · 2025: 4 · 2024: 4 · 2023: 1 · 2022: 4 · 2021: 4 · 2019: 3 · 2018: 1 · 2017: 8 · 2016: 2 · 2015: 4 · 2013: 7 · 2012: 4
FWCI
2.38
Type
journal article
Language
en
Open access
closed
Locations
  • Veterinary Parasitology (publishedVersion)
  • PubMed (publishedVersion)
  • CINECA IRIS Institutional Research Information System (University of Bari Aldo Moro) (submittedVersion)
Funders
University of Melbourne
Topics
  • Health Sciences › Medicine › Infectious Diseases › Parasitic Diseases Research and Treatment
  • Health Sciences › Veterinary › Small Animals › Helminth infection and control
  • Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Research on Leishmaniasis Studies
Keywords
Dirofilaria immitis, Dirofilaria repens, canine heartworm disease, filarial infection in mosquitoes, duplex real-time PCR, species differentiation, cytochrome c oxidase subunit I, ITS2, microfilariae, adult worms, Aedes albopictus, Aedes aegypti, Culex pipiens, molecular detection, disease surveillance, dogs
MeSH terms
Animals, Dirofilaria immitis, Dirofilariasis, Dog Diseases, Dogs, Culicidae, Species Specificity, Dirofilaria repens, Real-Time Polymerase Chain Reaction

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