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Archivos de medicina veterinaria

Print version ISSN 0301-732X

Abstract

FAGGION, SA et al. Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs. Arch. med. vet. [online]. 2013, vol.45, n.2, pp.197-201. ISSN 0301-732X.  http://dx.doi.org/10.4067/S0301-732X2013000200012.

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 cross-reactivity 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.

Keywords : canine monocytic ehrlichiosis; Ehrlichia canis; LAMP.

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