Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera Anaplasma and Ehrlichia in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies

• Published in: Pathogens (Basel) Vol. 10; no. 2; p. 192
• Main Authors: Sharma, Bhumika, R Ganta, Roman, Stone, Diana, Alhassan, Andy, Lanza-Perea, Marta, Matthew Belmar, Vanessa, Karasek, Inga, Cooksey, Elizabeth, M Butler, Catherine, Gibson, Kathryn, J Wilkerson, Melinda
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 10-02-2021; MDPI AG
• Subjects: Anaplasma; Ehrlichia; PCR; xMAP; Anaplasma; xMAP; Ehrlichia; PCR
• ISSN: 2076-0817; 2076-0817
• DOI: 10.3390/pathogens10020192

Infections with tick-borne pathogens belonging to
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
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).
infection in cattle had the highest detection rate (34.4%), followed by canines positive for
(16.4%) and
(13.9%). The assay aided in documenting the first molecular evidence for
in cattle and small ruminants and
and
in dogs in the Caribbean island of Grenada.