H 2 Detection and Species Identification of Bartonella Species by a Specific PCR Based Method

G. JOHNSON, S. RICHARSON, *R. TELLIER The Hospital for Sick Children, Toronto, Ontario

Objective: Several Bartonella species have been identified to date, most of which typically infect erythrocytes in their habitual host but may cause infection and disease in other hosts. Bartonella species implicated in human diseases include B. bacilliformis (Oroya fever); B. quintana (Trench Fever); B. henselae, which has been confirmed as the most common cause of cat scratch fever, with some cases being caused by B. quintana, B. clarridgeiae and possibly by Afipia felis; B. elizabethae and B. vinsonii subspecies berkhoffii, which have been reported in rare cases of bacteremia/endocarditis. Immunocompromised patients are susceptible to disseminated infection, such as in bacillary angiomatosis. Given the fastidiousness of  these agents, detection and identification by PCR would offer a rapid and sensitive diagnostic modality. 

Methods: A comprehensive PCR tests was designed, based on primers targeting a region of the riboflavin synthase  gene common to Bartonella species but distinct from other bacteria. 

Results: All the above Bartonella species could be detected by the PCR protocol. We determined the sequence of the amplicons for B. elizabethae and B. vinsonii subsp. berkhoffii to complete the reference database. Neither human DNA nor DNA from any bacterial species that was tested (including A. felis) was amplified in this assay. Species identification is achieved by digestion of the amplicon with a panel of restriction endonucleases. 

Conclusion: a comprehensive PCR assay for the detection and species identification of Bartonellae has been achieved and offers a rapid diagnostic test for these agents.