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Background   Bartonella  species are bacterial blood parasites of animals capable of causing disease in both animals and man. Cat-Scratch Disease (CSD) in humans is caused mainly by  Bartonella henselae  and is acquired from the cat, which serves as a reservoir for the bacteria. A second species,  B. clarridgeiae  is also implicated in the disease. Diagnosis of Bartonellosis by culture requires a week or more of incubation on enriched media containing blood, and recovery is often complicated by faster growing contaminating bacteria and fungi. PCR has been explored as an alternative to culture for both the detection and species identification of  Bartonella , however sensitivity problems have been reported and false negative reactions due to blood inhibitors have not generally been addressed in test design.    Methods  A novel, nested-PCR was designed for the detection of  Bartonella henselae  and  B. clarridgeiae  based on the strategy of targeting species-specific size differences in the 16S-23S rDNA intergenic regions. An Internal Amplification Control was used for detecting PCR inhibition. The nested-PCR was utilized in a study on 103 blood samples from pet and stray cats in Trinidad.    Results  None of the samples were positive by primary PCR, but the Nested-PCR detected  Bartonella  in 32/103 (31%) cats where 16 were infected with only  B. henselae , 13 with only  B. clarridgeiae  and 3 with both species. Of 22 stray cats housed at an animal shelter, 13 (59%) were positive for either or both species, supporting the reported increased incidence of  Bartonella  among feral cats.    Conclusion  The usefulness of a single PCR for the detection of  Bartonella henselae  and  B. clarridgeiae  in the blood of cats is questionable. A nested-PCR offers increased sensitivity over a primary PCR and should be evaluated with currently used methods for the routine detection and speciation of  Bartonella henselae  and  B. clarridgeiae . In Trinidad,  B. henselae  and  B. clarridgeiae  are the predominant species in cats and infection appears highest with stray cats, however  B. clarridgeiae  may be present at levels similar to that of  B. henselae  in the pet population.

bmc infectious diseases

A nested-PCR with an Internal Amplification Control for the detection and differentiation of Bartonella henselae and B. clarridgeiae: An examination of cats in Trinidad