High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms.
The canine hookworms Ancylostoma braziliense, Ancylostoma ceylanicum, Ancylostoma caninum and Uncinaria stenocephala are not only capable of producing morbidity and mortality in dogs but are also neglected tropical zoonoses. Each hookworm species differs considerably in its geographical distribution...
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ftdoajarticles:oai:doaj.org/article:bc2ab1f02e0c472d84e370cef3f2069e 2023-05-15T15:11:16+02:00 High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms. Luca Massetti Vito Colella Patsy A Zendejas Dinh Ng-Nguyen Lana Harriott Lara Marwedel Anke Wiethoelter Rebecca J Traub 2020-06-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0008392 https://doaj.org/article/bc2ab1f02e0c472d84e370cef3f2069e EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0008392 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008392 https://doaj.org/article/bc2ab1f02e0c472d84e370cef3f2069e PLoS Neglected Tropical Diseases, Vol 14, Iss 6, p e0008392 (2020) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2020 ftdoajarticles https://doi.org/10.1371/journal.pntd.0008392 2022-12-31T07:48:33Z The canine hookworms Ancylostoma braziliense, Ancylostoma ceylanicum, Ancylostoma caninum and Uncinaria stenocephala are not only capable of producing morbidity and mortality in dogs but are also neglected tropical zoonoses. Each hookworm species differs considerably in its geographical distribution, life cycle, biology, pathogenic impacts on both canine and human hosts, zoonotic potential, and response to treatment with anthelminthics. Here we describe the development and validation of two Taq-Man based multiplex PCR assays capable of detecting and differentiating all four canine hookworm species in faeces of naturally infected dogs. The analytical sensitivity of both assays was assessed using 10-fold serial dilutions of synthetic gene block fragments containing individual sequence targets of each hookworm species. The sensitivity of the assays and ability to detect mixed species infections were compared to a conventional PCR-Restriction Fragment Length Polymorphism based-approach when applied to laboratory and field samples from endemic areas. The qPCRs detected at least one species of hookworms in 82.4% of PCR-RFLP-negative but microscopy-positive samples. The qPCRs detected an additional 68% mixed infections with different species of canine hookworms, and additional single species infection with A. caninum (47%), U. stenocephala (33%) and A. ceylanicum (0.02%) that were missed by PCR-RFLP. These multiplex qPCR assays will assist field based epidemiological surveillance studies towards an accurate and sensitive monitoring of canine hookworm infections in dogs, to inform their species-specific zoonotic risks to populations living in endemic areas, globally. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 14 6 e0008392 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Luca Massetti Vito Colella Patsy A Zendejas Dinh Ng-Nguyen Lana Harriott Lara Marwedel Anke Wiethoelter Rebecca J Traub High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
The canine hookworms Ancylostoma braziliense, Ancylostoma ceylanicum, Ancylostoma caninum and Uncinaria stenocephala are not only capable of producing morbidity and mortality in dogs but are also neglected tropical zoonoses. Each hookworm species differs considerably in its geographical distribution, life cycle, biology, pathogenic impacts on both canine and human hosts, zoonotic potential, and response to treatment with anthelminthics. Here we describe the development and validation of two Taq-Man based multiplex PCR assays capable of detecting and differentiating all four canine hookworm species in faeces of naturally infected dogs. The analytical sensitivity of both assays was assessed using 10-fold serial dilutions of synthetic gene block fragments containing individual sequence targets of each hookworm species. The sensitivity of the assays and ability to detect mixed species infections were compared to a conventional PCR-Restriction Fragment Length Polymorphism based-approach when applied to laboratory and field samples from endemic areas. The qPCRs detected at least one species of hookworms in 82.4% of PCR-RFLP-negative but microscopy-positive samples. The qPCRs detected an additional 68% mixed infections with different species of canine hookworms, and additional single species infection with A. caninum (47%), U. stenocephala (33%) and A. ceylanicum (0.02%) that were missed by PCR-RFLP. These multiplex qPCR assays will assist field based epidemiological surveillance studies towards an accurate and sensitive monitoring of canine hookworm infections in dogs, to inform their species-specific zoonotic risks to populations living in endemic areas, globally. |
format |
Article in Journal/Newspaper |
author |
Luca Massetti Vito Colella Patsy A Zendejas Dinh Ng-Nguyen Lana Harriott Lara Marwedel Anke Wiethoelter Rebecca J Traub |
author_facet |
Luca Massetti Vito Colella Patsy A Zendejas Dinh Ng-Nguyen Lana Harriott Lara Marwedel Anke Wiethoelter Rebecca J Traub |
author_sort |
Luca Massetti |
title |
High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms. |
title_short |
High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms. |
title_full |
High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms. |
title_fullStr |
High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms. |
title_full_unstemmed |
High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms. |
title_sort |
high-throughput multiplex qpcrs for the surveillance of zoonotic species of canine hookworms. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2020 |
url |
https://doi.org/10.1371/journal.pntd.0008392 https://doaj.org/article/bc2ab1f02e0c472d84e370cef3f2069e |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 14, Iss 6, p e0008392 (2020) |
op_relation |
https://doi.org/10.1371/journal.pntd.0008392 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008392 https://doaj.org/article/bc2ab1f02e0c472d84e370cef3f2069e |
op_doi |
https://doi.org/10.1371/journal.pntd.0008392 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
14 |
container_issue |
6 |
container_start_page |
e0008392 |
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