Abstract
Infections with tick-borne pathogens belonging to Anaplasma/Ehrlichia 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 Anaplasma/Ehrlichia 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). Anaplasma marginale infection in cattle had the highest detection rate (34.4%), followed by canines positive for Anaplasma platys (16.4%) and Ehrlichia canis (13.9%). The assay aided in documenting the first molecular evidence for A. marginale in cattle and small ruminants and Ehrlichia chaffeensis and Ehrlichia ewingii in dogs in the Caribbean island of Grenada.
Highlights
For over three decades, Anaplasma and Ehrlichia species pathogens have been known to cause diseases in humans, while in pets and livestock these infections have been welldocumented for many decades [1,2,3]
The newly designed probe gave higher median fluorescence intensity (MFI) value for the xMAP assay when tested with the positive control plasmid
The final optimized xMAP protocol for all experiments was as follows: Polymerase chain reaction (PCR) was conducted for 35 cycles with an annealing at 50 ◦ C for 30 s, extension at 72 ◦ C for 30 s, and 2.5 mM
Summary
Anaplasma and Ehrlichia species pathogens have been known to cause diseases in humans, while in pets and livestock these infections have been welldocumented for many decades [1,2,3]. In the United States, human infections with Anaplasma and Ehrlichia species are identified as the second leading cause of tick-borne diseases after. Multi-infections with tickborne pathogens may enhance the disease severity and complicate the clinical presentation in a host [18,19,20]. Multiple-pathogen infections can persist for months to years and complicate a patient’s clinical presentation, substantially influencing the progression of the diseases, while creating challenges for laboratory diagnosis [13,22,23]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
