A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillance
Abstract Anthropogenic stressors are exacerbating the emergence and spread of pathogens worldwide. In regions like the Arctic, where ecosystems are particularly susceptible, marked changes are predicted in regional diversity, intensity, and patterns of infectious diseases. To understand such rapidly...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-11-01
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| Series: | Ecology and Evolution |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/ece3.10655 |
| _version_ | 1797448642982838272 |
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| author | Christina M. Tschritter Peter V. C. de Groot Marsha Branigan Markus Dyck Zhengxin Sun Stephen C. Lougheed |
| author_facet | Christina M. Tschritter Peter V. C. de Groot Marsha Branigan Markus Dyck Zhengxin Sun Stephen C. Lougheed |
| author_sort | Christina M. Tschritter |
| collection | DOAJ |
| description | Abstract Anthropogenic stressors are exacerbating the emergence and spread of pathogens worldwide. In regions like the Arctic, where ecosystems are particularly susceptible, marked changes are predicted in regional diversity, intensity, and patterns of infectious diseases. To understand such rapidly changing host‐pathogen dynamics and mitigate the impacts of novel pathogens, we need sensitive disease surveillance tools. We developed and validated a novel multiplexed, magnetic capture, and ddPCR tool for the surveillance of multiple pathogens in polar bears, a sentinel species that is considered susceptible to climate change and other stressors with a pan‐Arctic distribution. Through sequence‐specific magnetic capture, we concentrated five target template sequences from three zoonotic bacteria (Erysipelothrix rhusiopathiae, Francisella tularensis, and Mycobacterium tuberculosis complex) and two parasitic (Toxoplasma gondii and Trichinella spp.) pathogens from large quantities (<100 g) of host tissue. We then designed and validated two multiplexed probe‐based ddPCR assays for the amplification and detection of the low‐concentration target DNA. Validations used 48 polar bear tissues (muscle and liver). We detected 14, 1, 3, 4, and 22 tissue positives for E. rhusiopathiae, F. tularensis, M. tuberculosis complex, T. gondii, and Trichinella spp., respectively. These multiplexed assays offer a rapid, specific tool for quantifying and monitoring the changing geographical and host distributions of pathogens relevant to human and animal health. |
| first_indexed | 2024-03-09T14:13:22Z |
| format | Article |
| id | doaj.art-04fa418b97be4d8cb14049518b041484 |
| institution | Directory Open Access Journal |
| issn | 2045-7758 |
| language | English |
| last_indexed | 2024-03-09T14:13:22Z |
| publishDate | 2023-11-01 |
| publisher | Wiley |
| record_format | Article |
| series | Ecology and Evolution |
| spelling | doaj.art-04fa418b97be4d8cb14049518b0414842023-11-29T05:44:08ZengWileyEcology and Evolution2045-77582023-11-011311n/an/a10.1002/ece3.10655A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillanceChristina M. Tschritter0Peter V. C. de Groot1Marsha Branigan2Markus Dyck3Zhengxin Sun4Stephen C. Lougheed5Department of Biology Queen's University Kingston Ontario CanadaDepartment of Biology Queen's University Kingston Ontario CanadaDepartment of Environment and Natural Resources Government of the Northwest Territories Inuvik Northwest Territories CanadaDepartment of Environment Government of Nunavut Igloolik Nunavut CanadaDepartment of Biology Queen's University Kingston Ontario CanadaDepartment of Biology Queen's University Kingston Ontario CanadaAbstract Anthropogenic stressors are exacerbating the emergence and spread of pathogens worldwide. In regions like the Arctic, where ecosystems are particularly susceptible, marked changes are predicted in regional diversity, intensity, and patterns of infectious diseases. To understand such rapidly changing host‐pathogen dynamics and mitigate the impacts of novel pathogens, we need sensitive disease surveillance tools. We developed and validated a novel multiplexed, magnetic capture, and ddPCR tool for the surveillance of multiple pathogens in polar bears, a sentinel species that is considered susceptible to climate change and other stressors with a pan‐Arctic distribution. Through sequence‐specific magnetic capture, we concentrated five target template sequences from three zoonotic bacteria (Erysipelothrix rhusiopathiae, Francisella tularensis, and Mycobacterium tuberculosis complex) and two parasitic (Toxoplasma gondii and Trichinella spp.) pathogens from large quantities (<100 g) of host tissue. We then designed and validated two multiplexed probe‐based ddPCR assays for the amplification and detection of the low‐concentration target DNA. Validations used 48 polar bear tissues (muscle and liver). We detected 14, 1, 3, 4, and 22 tissue positives for E. rhusiopathiae, F. tularensis, M. tuberculosis complex, T. gondii, and Trichinella spp., respectively. These multiplexed assays offer a rapid, specific tool for quantifying and monitoring the changing geographical and host distributions of pathogens relevant to human and animal health.https://doi.org/10.1002/ece3.10655digital PCRdisease surveillancemultiplexUrsus maritimuszoonotic |
| spellingShingle | Christina M. Tschritter Peter V. C. de Groot Marsha Branigan Markus Dyck Zhengxin Sun Stephen C. Lougheed A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillance Ecology and Evolution digital PCR disease surveillance multiplex Ursus maritimus zoonotic |
| title | A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillance |
| title_full | A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillance |
| title_fullStr | A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillance |
| title_full_unstemmed | A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillance |
| title_short | A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillance |
| title_sort | new multiplexed magnetic capture droplet digital pcr tool for monitoring wildlife population health and pathogen surveillance |
| topic | digital PCR disease surveillance multiplex Ursus maritimus zoonotic |
| url | https://doi.org/10.1002/ece3.10655 |
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