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First isolation and molecular characterization of Ehrlichia canis in Costa Rica, Central America
Authors:LE Romero  AI Meneses  M Jiménez  DM Aguiar  G Dolz
Institution:a Laboratorio Central de Diagnóstico Veterinario, Ministerio de Agricultura y Ganadería (MAG), San Salvador, El Salvador
b Posgrado Regional en Ciencias Veterinarias Tropicales (PCVET), Escuela de Medicina Veterinaria, Universidad Nacional (UNA), Heredia, Costa Rica
c Laboratorio de Análisis Clínico, Escuela de Medicina Veterinaria, UNA, Heredia, Costa Rica
d Centro de Investigación en Hematología y Trastornos Afines (CIHATA), Universidad de Costa Rica (UCR), San José, Costa Rica
e Hospital Especies Menores, Escuela de Medicina Veterinaria, UNA, Heredia, Costa Rica
f Programa de Investigación en Medicina Poblacional (MEDPOB), Escuela de Medicina Veterinaria, UNA, Heredia, Costa Rica
g Departamento de Clinica Médica Veterinária, Faculdade de Agronomia e Medicina Veterinária da Universidade Federal de Mato Grosso, Cuiabá, MT, Brazil
h Departamento de Medicina Veterinária Preventiva e Saúde Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brazil
Abstract:The present study investigated Ehrlichia species in blood samples from dogs suspected of clinical ehrlichiosis, using molecular and isolation techniques in cell culture. From a total of 310 canine blood samples analyzed by 16S rRNA nested PCR, 148 (47.7%) were positive for Ehrlichia canis. DNA from Ehrlichia chaffeensis or Ehrlichia ewingii was not detected in any sample using species-specific primers in separated reactions. Leukocytes from five PCR-positive dogs were inoculated into DH82 cells; successful isolation of E. canis was obtained in four samples. Partial sequence of the dsb gene of eight canine blood samples (including the five samples for in vitro isolation) was obtained by PCR and their analyses through BLAST showed 100% of identity with the corresponding sequence of E. canis in GenBank. This study represents the first molecular diagnosis, isolation, and molecular characterization of E. canis in dogs from Costa Rica.
Keywords:Ehrlichia canis  Isolation  PCR  Costa Rica  Dogs
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