Abstract
BackgroundBartonella 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.MethodsA 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.ResultsNone 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.ConclusionThe 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.
Highlights
Bartonella species are bacterial blood parasites of animals capable of causing disease in both animals and man
We describe the development of a nested-Polymerase Chain Reaction (PCR) for the detection of B. henselae and B. clarridgeiae based on the strategy of species-specific size differences in the 16S-23S rDNA intergenic region that includes an Internal Amplification Control for PCR inhibitors
Of 103 blood samples subjected to PCR, none of the samples were positive for Bartonella in the primary reaction, but 32 were positive in the nested reaction
Summary
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. Bartonella are fastidious, gram-negative, bacteria comprised of at least 19 species and 3 subspecies [1] that are obligate parasites of the blood in reservoir animals. Studies suggest that other Bartonella species, known to cause disease in humans, are found in the cat Bartonella henselae and, to a lesser extent, Bartonella clarridgeiae are known to cause Cat-Scratch Disease (CSD) in humans [8]; see [9] for review of CSD
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