Assessment of a quantitative 5' nuclease real-time polymerase chain reaction using the nicotinamide adenine dinucleotide dehydrogenase gamma subunit (nuoG) for Bartonella species in domiciled and stray cats in Brazil
Nenhuma Miniatura disponível
Data
2015
Autores
André, Marcos Rogério [UNESP]
Dumler, John Stephen
Herrera, Heitor M.
Gonçalves, Luiz R. [UNESP]
Sousa, Keyla C. M. de [UNESP]
Scorpio, Diana Gerardi
Santis, Ana Cláudia Gabriela Alexandre de [UNESP]
Domingos, Iara Helena
Macedo, Gabriel Carvalho de
Machado, Rosangela Zacarias [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
SAGE Publications
Resumo
The objective of this study was to develop a quantitative 5' nuclease real-time polymerase chain reaction (PCR) assay to diagnose infections caused by Bartonella species. Between January and April 2013 whole blood samples were collected by convenience from 151 cats (86 domiciled and 65 stray cats). The feline blood samples were subjected to a novel quantitative 5' nuclease real-time PCR (qPCR) for Bartonella species targeting the nictonamide adenine dinucleotide dehydrogenase gamma subunit (nuoG) and conventional PCR assays targeting intergenic transcribed spacer, ribC, gltA, pap31 and rpoB, followed by sequencing and basic local alignment search tool analysis. The qPCR assay detected as few as 10 copies of plasmid per reaction. Forty-six (54.4% domiciled and 45.6% stray cats) of 151 sampled cats showed positive results in nuoG qPCR for Bartonella species. The absolute quantification of nuoG Bartonella DNA in sampled cats ranged from 1.1 × 10(4) to 1.3 × 10(4). Eighteen (39.1%) of 46 positive samples in the qPCR were also positive in conventional PCR assays. The sequencing confirmed that Bartonella henselae and Bartonella clarridgeiae circulate in cats in midwestern Brazil. The present work provides details of a novel qPCR assay to diagnose infections caused by Bartonella species.
Descrição
Palavras-chave
Como citar
Journal Of Feline Medicine And Surgery, p. 1-9, 2015.