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Specific In Situ Hybridization of Haemobartonella felis with a DNA Probe and Tyramide Signal Amplification

Department of Veterinary Pathobiology, University of Illinois, Urbana, IL (LMB, JBM, SKC); and Division of Drug Safety Evaluation, Abbott Laboratories, Abbott Park, IL (PKC)

Haemobartonella felis is an epierythrocytic bacterium suspected to be the causative agent of feline infectious anemia. Previous studies with a polymerase chain reaction assay have identified a mycoplasmal 16S rRNA gene sequence that coincides with clinical disease and the presence of organisms in the blood. Tissues from a cat experimentally infected with H. felis were used for in situ hybridization studies to physically link this 16S rRNA gene to the organisms on the red cells. A biotin-labeled probe was used in conjunction with tyramide signal amplification to visualize the hybridization signal. This study clearly demonstrates a specific hybridization signal on the red cells in the tissues of the H. felis-infected cat. This in situ hybridization study is the final step in fulfilling the molecular guidelines for disease causation and proves that H. felis, a mycoplasmal organism, is the causative agent of feline infectious anemia.

Key words: feline; Haemobartonella felis; in situ hybridization; Mycoplasma; TSA.

Request reprints from Dr. Linda M. Berent, 2741 VMBSB, Department of Veterinary Pathobiology, 2001 S. Lincoln Avenue, Urbana, IL 61802 (USA).

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