title: [Real-time PCR-based detection of Bartonella vinsonii sub sp. berkhoffii by TaqMan minor groove binder probe].
abstract: OBJECTIVE
  Bartonella vinsonii subsp. berkhoffii is a fastidious haemotropic Gram-negative bacterium that has been identified as an emerging causative agent for zoonotic diseases of human and dogs. This study aimed to develop a TaqMan-MGB probe based, highly sensitive and species-specific fluorescence quantitative PCR assay for rapid detection of B. vinsonii subsp. berkhoffii.
  
  
  METHODS
  The species-specific primersand probe set for B. vinsonii subsp. berkhoffii were designed. The annealing temperature, final concentration of the TaqMan-MGB probe and primers were optimized. Specificity, sensitivity and reproducibility of the PCR system were assessed. The standard curve was made using 10 x dilution series of the plasmid standard to analyze stability and PCR efficiency.
  
  
  RESULTS
  The real-time PCR with TaqMan-MGB assay was highly specific and sensitive for the detection of B. vinsonii subsp. berkhoffii. TaqMan-MGB probe-based fluorescence quantitative PCR did not show cross reactivity with the other Bartonella species, non-Bartonella bacteria and dogs and human. The detection limit of the TaqMan-MGB assay for the detection of B. vinsonii subsp. berkhoffii was 11 copies of the plasmid DNA per PCR reaction. The coefficient of variation CV% from the intra- and inter-group was in the range of 0.12% -0.70% and 0.14-0.55% which were acceptable. The correlation coefficient and E-value of the standard curve were 1.0 and 104.7%, which reflected a very good linearity and high efficiency.
  
  
  CONCLUSIONS
  The TaqMan MGB-based probe fluorescence quantitative PCR assay was a reliable, species-specific, sensitive and useful tool for rapid detection of B. vinsonii subsp. berkhoffii.
authors.affiliation: Department of Vector Biology and Control, State Key Laboratory for Infectious Diseases Prevention and Control, WHO Collaborating Centre for Vector Surveillance and Management, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China. lidongmei@icdc.cn
authors.forename: Dongmei
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Vector Biology and Control, State Key Laboratory for Infectious Diseases Prevention and Control, WHO Collaborating Centre for Vector Surveillance and Management, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Xiuping
authors.initials: X
authors.lastname: Song
authors.affiliation: Department of Vector Biology and Control, State Key Laboratory for Infectious Diseases Prevention and Control, WHO Collaborating Centre for Vector Surveillance and Management, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Vector Biology and Control, State Key Laboratory for Infectious Diseases Prevention and Control, WHO Collaborating Centre for Vector Surveillance and Management, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Qiyong
authors.initials: Q
authors.lastname: Liu
mesh_terms: D000818:Animals; D001473:Bartonella / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001474:Bartonella Infections / Q000175:diagnosis / Q000382:microbiology* / Q000662:veterinary*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004283:Dog Diseases / Q000175:diagnosis / Q000382:microbiology*; D004285:Dogs; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000737:chemistry / Q000235:genetics; D015047:Zoonoses / Q000175:diagnosis / Q000382:microbiology
chemical_list: D015342:DNA Probes; D019914:Taq Polymerase