Molecular Detection of spp. in China and St. Kitts.

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