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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Development and evaluation of a loop-mediated isothermal amplification assay for detection of Ehrlichia canis DNA in naturally infected dogs using the p30 gene

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Author(s):
Pinhanelli, V. C. [1] ; Costa, P. N. M. [1] ; Silva, G. [1] ; Aguiar, D. M. [2] ; Silva, C. M. L. [1] ; Fachin, A. L. [1] ; Marins, M. [1]
Total Authors: 7
Affiliation:
[1] Univ Ribeirao Preto, Unidade Biotecnol, Ribeirao Preto, SP - Brazil
[2] Univ Fed Mato Grosso, Fac Agron Med Vet & Zootecnia, Cuiaba, MT - Brazil
Total Affiliations: 2
Document type: Journal article
Source: Genetics and Molecular Research; v. 14, n. 4, p. 17885-17892, 2015.
Web of Science Citations: 0
Abstract

Canine monocytic ehrlichiosis (CME) is a common tick-borne disease caused by the rickettsial bacterium Ehrlichia canis (Rickettsiales: Anaplasmataceae). In view of the different stages and variable clinical signs of CME, which can overlap with those of other infections, a conclusive diagnosis can more readily be obtained by combining clinical and hematological evaluations with molecular diagnostic methods. In this study, a loop-mediated isothermal amplification (LAMP) assay targeting the p30 gene of E. canis was developed. The assay was developed using DNA extracted from E. canis-infected cultures of the macrophage cell line DH82 and samples from dogs testing positive for E. canis DNA by PCR. The LAMP assay was compared to a p30-based PCR assay, using DNA extracted from EDTA-anticoagulated blood samples of 137 dogs from an endemic region in Brazil. The LAMP assay was sensitive enough to detect a single copy of the target gene, and identified 74 (54.0%) E. canis DNA-positive samples, while the p30 PCR assay detected 50 positive samples (36.5%) among the field samples. Agreement between the two assays was observed in 42 positive and 55 negative samples. However, 32 positive samples that were not detected by the PCR assay were identified by the LAMP assay, while eight samples identified as E. canis-positive by PCR showed negative results in LAMP. The developed E. canis LAMP assay showed the potential to maximize the use of nucleic acid tests in a veterinary clinical laboratory, and to improve the diagnosis of CME. (AU)

FAPESP's process: 12/15940-6 - Delopment and evaluation of an assay based in the loop mediated isothermal amplification of DNA (lamp) technique for the detection of Ehrlichia canis in naturally infected dogs
Grantee:Vitor Caressato Pinhanelli
Support Opportunities: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 12/01235-9 - Delopment and evaluation of an assay based in the loop mediated isothermal amplification of DNA (LAMP) techqnique for the detection of Ehrlichia canis in naturally infected dogs
Grantee:Mozart de Azevedo Marins
Support Opportunities: Regular Research Grants