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

作者：Andre Marcos Rogerio^*; Dumler John Stephen; Herrera Heitor M; Goncalves Luiz R; de Sousa Keyla C M; Scorpio Diana Gerardi; Gabriela Alexandre de Santis Ana Claudia; Domingos Iara Helena; de Macedo Gabriel Carvalho; Machado Rosangela Zacarias

来源：Journal of Feline Medicine and Surgery, 2016, 18(10): 783-790.

DOI：10.1177/1098612X15593787

Objectives 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.
Methods 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.
Results 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 x 10(4) to 1.3 x 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.
Conclusions and relevance The present work provides details of a novel qPCR assay to diagnose infections caused by Bartonella species.

• 出版日期2016-10