Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific
The Asia-Pacific hosts a large diversity of canine vector-borne pathogens (VBPs) with some of the most common and most pathogenic, generating significant mortality as well as a spectrum of health impacts on local dog populations. The VBPs <i>Anaplasma platys</i>, <i>Babesia gibsoni...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-05-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/9/5/1092 |
| _version_ | 1797533492526972928 |
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| author | Lucas Huggins Luca Massetti Bettina Schunack Vito Colella Rebecca Traub |
| author_facet | Lucas Huggins Luca Massetti Bettina Schunack Vito Colella Rebecca Traub |
| author_sort | Lucas Huggins |
| collection | DOAJ |
| description | The Asia-Pacific hosts a large diversity of canine vector-borne pathogens (VBPs) with some of the most common and most pathogenic, generating significant mortality as well as a spectrum of health impacts on local dog populations. The VBPs <i>Anaplasma platys</i>, <i>Babesia gibsoni</i>, <i>Babesia vogeli</i>, <i>Ehrlichia canis</i>, <i>Hepatozoon canis</i> and haemotropic <i>Mycoplasma</i> spp. are all endemic throughout the region, with many exhibiting shifting geographical distributions that warrant urgent attention. Moreover, many of these species cause similar clinical signs when parasitising canine hosts, whilst knowledge of the exact pathogen is critical to ensure treatment is effective. This is complicated by frequent coinfection that can exacerbate pathology. Here, we describe the development, optimisation and validation of two novel quadruplex Taq-Man based real-time PCRs (qPCRs) for the specific and sensitive detection of the aforementioned VBPs. To ensure accurate evaluation of diagnostic performance, results of our qPCRs were evaluated on field samples from Thai dogs and compared with both conventional PCR (cPCR) results and next-generation sequencing (NGS) metabarcoding. Our qPCRs were found to be more sensitive at detecting canine VBP than cPCR and generated results similar to those achieved by NGS. These qPCRs will provide a valuable high-throughput diagnostic tool available to epidemiologists, researchers and clinicians for the diagnosis of key canine VBPs in the Asia-Pacific and further afield. |
| first_indexed | 2024-03-10T11:15:18Z |
| format | Article |
| id | doaj.art-893a527b985047928ba525e4e62670b7 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T11:15:18Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-893a527b985047928ba525e4e62670b72023-11-21T20:28:13ZengMDPI AGMicroorganisms2076-26072021-05-0195109210.3390/microorganisms9051092Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-PacificLucas Huggins0Luca Massetti1Bettina Schunack2Vito Colella3Rebecca Traub4Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, VIC 3052, AustraliaFaculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, VIC 3052, AustraliaElanco GmbH, Heinz-Lohmann-Str. 4, 27472 Cuxhaven, GermanyFaculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, VIC 3052, AustraliaFaculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, VIC 3052, AustraliaThe Asia-Pacific hosts a large diversity of canine vector-borne pathogens (VBPs) with some of the most common and most pathogenic, generating significant mortality as well as a spectrum of health impacts on local dog populations. The VBPs <i>Anaplasma platys</i>, <i>Babesia gibsoni</i>, <i>Babesia vogeli</i>, <i>Ehrlichia canis</i>, <i>Hepatozoon canis</i> and haemotropic <i>Mycoplasma</i> spp. are all endemic throughout the region, with many exhibiting shifting geographical distributions that warrant urgent attention. Moreover, many of these species cause similar clinical signs when parasitising canine hosts, whilst knowledge of the exact pathogen is critical to ensure treatment is effective. This is complicated by frequent coinfection that can exacerbate pathology. Here, we describe the development, optimisation and validation of two novel quadruplex Taq-Man based real-time PCRs (qPCRs) for the specific and sensitive detection of the aforementioned VBPs. To ensure accurate evaluation of diagnostic performance, results of our qPCRs were evaluated on field samples from Thai dogs and compared with both conventional PCR (cPCR) results and next-generation sequencing (NGS) metabarcoding. Our qPCRs were found to be more sensitive at detecting canine VBP than cPCR and generated results similar to those achieved by NGS. These qPCRs will provide a valuable high-throughput diagnostic tool available to epidemiologists, researchers and clinicians for the diagnosis of key canine VBPs in the Asia-Pacific and further afield.https://www.mdpi.com/2076-2607/9/5/1092multiplex qPCRcanine vector-borne diseasedogsmolecular diagnosticsAsia-Pacific<i>Anaplasma</i> <i>platys</i> |
| spellingShingle | Lucas Huggins Luca Massetti Bettina Schunack Vito Colella Rebecca Traub Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific Microorganisms multiplex qPCR canine vector-borne disease dogs molecular diagnostics Asia-Pacific <i>Anaplasma</i> <i>platys</i> |
| title | Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific |
| title_full | Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific |
| title_fullStr | Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific |
| title_full_unstemmed | Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific |
| title_short | Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific |
| title_sort | novel high throughput multiplex qpcrs for the detection of canine vector borne pathogens in the asia pacific |
| topic | multiplex qPCR canine vector-borne disease dogs molecular diagnostics Asia-Pacific <i>Anaplasma</i> <i>platys</i> |
| url | https://www.mdpi.com/2076-2607/9/5/1092 |
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