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https://www.arca.fiocruz.br/handle/icict/64705
COMPARATIVE STUDY OF COLORIMETRIC IN SITU HYBRIDIZATION AND QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION FOR DIAGNOSIS OF INFECTION BY LEISHMANIA INFANTUM IN DOGS IN FORMALIN-FIXED AND PARAFFIN-EMBEDDED SKIN
Author
Silva, Ricardo Gonçalves
Kiupel, Matti
Langohr, Ingeborg Maria
Wise, Annabel
Pereira, Sandro Antonio
Gonçalves, Natália Pedra
Conceição, Greice Maria Silva da
Ferreira, Luiz Cláudio
Campos, Monique Paiva de
Miranda, Luciana de Freitas Campos
Figueiredo, Fabiano Borges
Oliveira, Raquel de Vasconcellos Carvalhaes de
Keidel, Lucas
Menezes, Rodrigo Caldas
Kiupel, Matti
Langohr, Ingeborg Maria
Wise, Annabel
Pereira, Sandro Antonio
Gonçalves, Natália Pedra
Conceição, Greice Maria Silva da
Ferreira, Luiz Cláudio
Campos, Monique Paiva de
Miranda, Luciana de Freitas Campos
Figueiredo, Fabiano Borges
Oliveira, Raquel de Vasconcellos Carvalhaes de
Keidel, Lucas
Menezes, Rodrigo Caldas
Affilliation
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Michigan State University. Veterinary Diagnostic Laboratory. Lansing, MI, USA.
Sanofi. Global Discovery Pathology. Translational Models Research Platform. Cambridge, MA, USA.
Michigan State University. Veterinary Diagnostic Laboratory. Lansing, MI, USA.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Institute of Technology in Immunobiologicals (Bio-Manguinhos). Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Institute of Technology in Immunobiologicals (Bio-Manguinhos). Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Carlos Chagas Institute. Curitiba, PR, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research and Surveillance of Leishmaniasis. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Carlos Chagas Institute. Curitiba, PR, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Epidemiology. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Michigan State University. Veterinary Diagnostic Laboratory. Lansing, MI, USA.
Sanofi. Global Discovery Pathology. Translational Models Research Platform. Cambridge, MA, USA.
Michigan State University. Veterinary Diagnostic Laboratory. Lansing, MI, USA.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Institute of Technology in Immunobiologicals (Bio-Manguinhos). Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Institute of Technology in Immunobiologicals (Bio-Manguinhos). Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Carlos Chagas Institute. Curitiba, PR, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research and Surveillance of Leishmaniasis. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Carlos Chagas Institute. Curitiba, PR, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Epidemiology. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. Evandro Chagas National Institute of Infectious Diseases. Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals. Rio de Janeiro, RJ, Brazil.
Abstract
The zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and dogs are reservoirs for this parasite. For the diagnosis of Leishmania at the species level in dogs in formalin-fixed, paraffin-embedded skin (FFPES) samples, colorimetric in situ hybridization (CISH) and quantitative real-time polymerase chain reaction (qPCR) are options, but their sensitivities are not well established. Therefore, the aim of this study was to determine the sensitivity of these two techniques in FFPES for the diagnosis of the L. infantum infection in dogs using culture as the reference standard. The FFPES of 48 dogs with cutaneous infection by L. infantum confirmed by culture and by multilocus enzyme electrophoresis were examined by CISH and qPCR using specific probes for L. infantum. The sensitivities of qPCR, CISH and their combination were, respectively, 77.0%, 58.0% and 83.3%. The sensitivities of qPCR in dogs with and without clinical signs were, respectively, 74.2% and 82.4%. The sensitivities of CISH in dogs with and without clinical signs were, respectively, 61.3% and 52.9%. The CISH and qPCR showed satisfactory sensitivities for the diagnosis of L. infantum in the FFPES of dogs, even in dogs without clinical signs, and their combination increases the sensitivity for this diagnosis.
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