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Bartonella  are vector-borne hemotropic bacteria that infect a wide variety of hosts, including people. While there are PCR assays that can identify individual or groups of  Bartonella , there is no reliable molecular method to simultaneously detect all species while maintaining genus specificity and sensitivity. By comparing highly conserved  16S rRNA  sequences of the better-recognized  Bartonella  spp. on GenBank, we selected primers and probes for a genus-specific pan- Bartonella  FRET-qPCR. Then, a  gltA -based  Bartonella  PCR was established by selecting primers for a highly variable region of  gltA , of which the sequenced amplicons could identify individual  Bartonella  spp. The pan- Bartonella  FRET-qPCR did not detect negative controls ( Brucella  spp.,  Anaplasma  spp.,  Rickettsia  spp.,  Coxiella burnetii , and  Wolbachia ) but reliably detected as few as two copies of the positive control ( Bartonella henselae ) per reaction. There was complete agreement between the pan- Bartonella  FRET-qPCR and the  gltA -based  Bartonella  PCR in detecting  Bartonella  in convenience test samples from China and St. Kitts: cats (26%; 81/310),  Ctenocephalides felis  (20%; 12/60), cattle (24%; 23/98), and donkeys (4%; 1/20). Sequencing of the  gltA -based  Bartonella  PCR products revealed  B. henselae  (70%; 57/81) and  B. clarridgeiae  (30%; 24/81) in cats and  C. felis  (67%; 8/12, and 33%; 4/12, respectively) and  B. bovis  in cattle (23.5%; 23/98) and donkeys (4.0%; 1/24). The pan- Bartonella  FRET-qPCR and  gltA -based  Bartonella  PCR we developed are highly sensitive and specific in detecting recognized  Bartonella  spp. in a single reaction. The pan- Bartonella  FRET-qPCR is convenient requiring no gel electrophoresis and providing copy numbers, while the  gltA -based  Bartonella  PCR reliably differentiates individual  Bartonella  species. The use of these PCRs should greatly facilitate large-scale surveillance studies and the diagnosis of infections in clinical samples.

Molecular Detection ofBartonellaspp. in China and St. Kitts