Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera <i>Anaplasma</i> and <i>Ehrlichia</i> in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies
Infections with tick-borne pathogens belonging to <i>Anaplasma/Ehrlichia</i> in various vertebrate hosts are a persistent problem resulting in nonspecific clinical signs during early infection. Diagnosis of single and multi-infections with these pathogens, causing diseases in companion/a...
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| Format: | Article |
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MDPI AG
2021-02-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/2/192 |
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| author | Bhumika Sharma Roman R. Ganta Diana Stone Andy Alhassan Marta Lanza-Perea Vanessa Matthew Belmar Inga Karasek Elizabeth Cooksey Catherine M. Butler Kathryn Gibson Melinda J. Wilkerson |
| author_facet | Bhumika Sharma Roman R. Ganta Diana Stone Andy Alhassan Marta Lanza-Perea Vanessa Matthew Belmar Inga Karasek Elizabeth Cooksey Catherine M. Butler Kathryn Gibson Melinda J. Wilkerson |
| author_sort | Bhumika Sharma |
| collection | DOAJ |
| description | Infections with tick-borne pathogens belonging to <i>Anaplasma/Ehrlichia</i> in various vertebrate hosts are a persistent problem resulting in nonspecific clinical signs during early infection. Diagnosis of single and multi-infections with these pathogens, causing diseases in companion/agricultural animals and people, remains a challenge. Traditional methods of diagnosis, such as microscopy and serology, have low sensitivity and specificity. Polymerase chain reaction (PCR) assays are widely used to detect early-phase infections, since these have high sensitivity and specificity. We report the development and validation of an assay involving PCR followed by magnetic capture method using species-specific oligonucleotides to detect six <i>Anaplasma/Ehrlichia</i> species pathogens in canine, bovine, caprine, and ovine blood samples. Overall, the assay application to 455 samples detected 30.1% (137/455) positives for one or more out of six screened pathogens. Single-pathogen infections were observed in 94.9% (130/137) of the positive samples, while co-infections were detected in 5.1% (7/137). <i>Anaplasma marginale</i> infection in cattle had the highest detection rate (34.4%), followed by canines positive for <i>Anaplasma platys</i> (16.4%) and <i>Ehrlichia canis</i> (13.9%). The assay aided in documenting the first molecular evidence for <i>A. marginale</i> in cattle and small ruminants and <i>Ehrlichia chaffeensis</i> and <i>Ehrlichia ewingii</i> in dogs in the Caribbean island of Grenada. |
| first_indexed | 2024-03-09T04:51:26Z |
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| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-09T04:51:26Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-e951e5b639394641b0d33078dfb1aa2b2023-12-03T13:10:41ZengMDPI AGPathogens2076-08172021-02-0110219210.3390/pathogens10020192Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera <i>Anaplasma</i> and <i>Ehrlichia</i> in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West IndiesBhumika Sharma0Roman R. Ganta1Diana Stone2Andy Alhassan3Marta Lanza-Perea4Vanessa Matthew Belmar5Inga Karasek6Elizabeth Cooksey7Catherine M. Butler8Kathryn Gibson9Melinda J. Wilkerson10Department of Pathobiology, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaCenter of Excellence of Vector Borne Diseases, Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, USADepartment of Pathobiology, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaDepartment of Pathobiology, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaDepartment of Small Animal Medicine & Surgery, School of Veterinary Medicine, St Georges University, St. George, West Indies, GrenadaDepartment of Pathobiology, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaDepartment of Large Animal Medicine & Surgery, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaDepartment of Large Animal Medicine & Surgery, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaDepartment of Large Animal Medicine & Surgery, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaDepartment of Pathobiology, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaDepartment of Pathobiology, School of Veterinary Medicine, St. George’s University, St. George, West Indies, GrenadaInfections with tick-borne pathogens belonging to <i>Anaplasma/Ehrlichia</i> in various vertebrate hosts are a persistent problem resulting in nonspecific clinical signs during early infection. Diagnosis of single and multi-infections with these pathogens, causing diseases in companion/agricultural animals and people, remains a challenge. Traditional methods of diagnosis, such as microscopy and serology, have low sensitivity and specificity. Polymerase chain reaction (PCR) assays are widely used to detect early-phase infections, since these have high sensitivity and specificity. We report the development and validation of an assay involving PCR followed by magnetic capture method using species-specific oligonucleotides to detect six <i>Anaplasma/Ehrlichia</i> species pathogens in canine, bovine, caprine, and ovine blood samples. Overall, the assay application to 455 samples detected 30.1% (137/455) positives for one or more out of six screened pathogens. Single-pathogen infections were observed in 94.9% (130/137) of the positive samples, while co-infections were detected in 5.1% (7/137). <i>Anaplasma marginale</i> infection in cattle had the highest detection rate (34.4%), followed by canines positive for <i>Anaplasma platys</i> (16.4%) and <i>Ehrlichia canis</i> (13.9%). The assay aided in documenting the first molecular evidence for <i>A. marginale</i> in cattle and small ruminants and <i>Ehrlichia chaffeensis</i> and <i>Ehrlichia ewingii</i> in dogs in the Caribbean island of Grenada.https://www.mdpi.com/2076-0817/10/2/192<i>Anaplasma</i><i>Ehrlichia</i>PCRxMAP |
| spellingShingle | Bhumika Sharma Roman R. Ganta Diana Stone Andy Alhassan Marta Lanza-Perea Vanessa Matthew Belmar Inga Karasek Elizabeth Cooksey Catherine M. Butler Kathryn Gibson Melinda J. Wilkerson Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera <i>Anaplasma</i> and <i>Ehrlichia</i> in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies Pathogens <i>Anaplasma</i> <i>Ehrlichia</i> PCR xMAP |
| title | Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera <i>Anaplasma</i> and <i>Ehrlichia</i> in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies |
| title_full | Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera <i>Anaplasma</i> and <i>Ehrlichia</i> in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies |
| title_fullStr | Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera <i>Anaplasma</i> and <i>Ehrlichia</i> in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies |
| title_full_unstemmed | Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera <i>Anaplasma</i> and <i>Ehrlichia</i> in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies |
| title_short | Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera <i>Anaplasma</i> and <i>Ehrlichia</i> in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies |
| title_sort | development of a multiplex pcr and magnetic dna capture assay for detecting six species pathogens of the genera i anaplasma i and i ehrlichia i in canine bovine caprine and ovine blood samples from grenada west indies |
| topic | <i>Anaplasma</i> <i>Ehrlichia</i> PCR xMAP |
| url | https://www.mdpi.com/2076-0817/10/2/192 |
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