title: 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.
abstract: 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.
authors.affiliation: Paulista State University (UNESP), Jaboticabal, São Paulo, Brazil marcos_andre@fcav.unesp.br.
authors.forename: Marcos Rogério
authors.initials: MR
authors.lastname: André
authors.affiliation: University of Maryland, Baltimore, MD, USA Johns Hopkins School of Medicine, Baltimore, MD, USA.
authors.forename: John Stephen
authors.initials: JS
authors.lastname: Dumler
authors.affiliation: Dom Bosco Catholic University, Campo Grande, Mato Grosso do Sul, Brazil.
authors.forename: Heitor M
authors.initials: HM
authors.lastname: Herrera
authors.affiliation: Paulista State University (UNESP), Jaboticabal, São Paulo, Brazil.
authors.forename: Luiz R
authors.initials: LR
authors.lastname: Gonçalves
authors.affiliation: Paulista State University (UNESP), Jaboticabal, São Paulo, Brazil.
authors.forename: Keyla Cm
authors.initials: KC
authors.lastname: de Sousa
authors.affiliation: Ross University School of Veterinary Medicine, Basseterre, St Kitts, West Indies.
authors.forename: Diana Gerardi
authors.initials: DG
authors.lastname: Scorpio
authors.affiliation: Paulista State University (UNESP), Jaboticabal, São Paulo, Brazil.
authors.forename: Ana Cláudia Gabriela Alexandre
authors.initials: AC
authors.lastname: de Santis
authors.affiliation: Zoonosis Control Center (CCZ), Campo Grande, Mato Grosso do Sul, Brazil.
authors.forename: Iara Helena
authors.initials: IH
authors.lastname: Domingos
authors.affiliation: Dom Bosco Catholic University, Campo Grande, Mato Grosso do Sul, Brazil.
authors.forename: Gabriel Carvalho
authors.initials: GC
authors.lastname: de Macedo
authors.affiliation: Paulista State University (UNESP), Jaboticabal, São Paulo, Brazil.
authors.forename: Rosangela Zacarias
authors.initials: RZ
authors.lastname: Machado
mesh_terms: D000818:Animals; D000835:Animals, Wild; D001473:Bartonella / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001474:Bartonella Infections / Q000175:diagnosis / Q000662:veterinary*; D001938:Brazil / Q000453:epidemiology; D002371:Cat Diseases / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D002415:Cats; D004269:DNA, Bacterial / Q000032:analysis; D005260:Female; D006799:Housing, Animal; D008297:Male; D009243:NAD / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D009243:NAD