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- Title
Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs.
- Authors
Faggion, S. A.; Salvador, A. R.; Jacobino, K. L.; Bortolotto, L. F. B.; Lopes, M. B.; Silva, M.; Santos, E. V.; Fachin, A. L.; França, S. C.; Marins, M .
- Abstract
The rickettsial bacterium Ehrlichia canis is the etiological agent of canine monocytic ehrlichiosis, one of the most important canine tick-borne diseases in the world. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for detection of E. canis DNA using LAMP primers targeting the groESL operon. Reactions were performed at 60°C for 60 min and the results were visualized by gel electrophoresis. Successful amplification was obtained using plasmid DNA containing a fragment of the groESL operon and DNA extracted from blood samples that tested positive for E. canis by real-time PCR. The specificity of amplification was confirmed by EcoRI restriction of internal sites in the LAMP primers and no crossreactivity with blood samples positive for Babesia spp., another common tick-borne pathogen, was observed. The high cost of nucleic acid tests (NAT) is one of the disadvantages for their large-scale use as routine diagnostic tests. The E. canis LAMP assay developed here is an interesting alternative to PCR since it does not require a thermocycler, thus reducing costs for the veterinary clinical laboratory.
- Subjects
GENE amplification; EHRLICHIA; BLOOD sampling; DNA analysis; DOGS; ANIMAL health; RICKETTSIALES
- Publication
Archivos de Medicina Veterinaria, 2013, Vol 45, Issue 2, p197
- ISSN
0301-732X
- Publication type
Article
- DOI
10.4067/S0301-732X2013000200012