Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs

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...

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Autores principales:	Faggion,SA, Salvador,AR, Jacobino,KL, Bortolotto,LFB, Lopes,MB, Silva,M, Santos,EV, Fachin,AL, França,SC, Marins,M
Lenguaje:	English
Publicado:	Facultad de Ciencias Veterinarias, Universidad Austral de Chile 2013
Materias:	canine monocytic ehrlichiosis Ehrlichia canis LAMP
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0301-732X2013000200012
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spelling	oai:scielo:S0301-732X20130002000122013-09-03Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogsFaggion,SASalvador,ARJacobino,KLBortolotto,LFBLopes,MBSilva,MSantos,EVFachin,ALFrança,SCMarins,M canine monocytic ehrlichiosis Ehrlichia canis LAMP 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.info:eu-repo/semantics/openAccessFacultad de Ciencias Veterinarias, Universidad Austral de ChileArchivos de medicina veterinaria v.45 n.2 20132013-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0301-732X2013000200012en10.4067/S0301-732X2013000200012
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	canine monocytic ehrlichiosis Ehrlichia canis LAMP
spellingShingle	canine monocytic ehrlichiosis Ehrlichia canis LAMP Faggion,SA Salvador,AR Jacobino,KL Bortolotto,LFB Lopes,MB Silva,M Santos,EV Fachin,AL França,SC Marins,M Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs
description	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.
author	Faggion,SA Salvador,AR Jacobino,KL Bortolotto,LFB Lopes,MB Silva,M Santos,EV Fachin,AL França,SC Marins,M
author_facet	Faggion,SA Salvador,AR Jacobino,KL Bortolotto,LFB Lopes,MB Silva,M Santos,EV Fachin,AL França,SC Marins,M
author_sort	Faggion,SA
title	Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs
title_short	Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs
title_full	Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs
title_fullStr	Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs
title_full_unstemmed	Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs
title_sort	loop-mediated isothermal amplification assay for the detection of ehrlichia canis dna in blood samples from dogs
publisher	Facultad de Ciencias Veterinarias, Universidad Austral de Chile
publishDate	2013
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0301-732X2013000200012
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Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs

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