THE DEVELOPMENT OF AN ENZYME-LINKED IMMUNOSORBENT FOR THE DETECTION OF BARTONELLA HENSELAE INFECTION IN CATS

Abstract

The study was conducted to determine the prevalence of Bartonella spp. in two hundred and fifty well-care client-owned cats in Bangkok metropolitan area between November 2010-November 2011. These cats had no clinical signs at the time of presentation and no visual flea infestation. Blood collection was performed and cultured in 5% sheep blood agar. The species of Bartonella spp. was identified with Polymerase Chain Reaction (PCR) and gene sequencing. This study indicated that the prevalence of Bartonella spp. in well-care cats in Bangkok metropolitan was 4.4% (11/250) with Bartonella henselae 91% (10/11) and Bartonella claridgeiae 9% (1/11). DNA of B. henselae was used as a DNA template for Polymerase Chain Reaction (PCR) targeting 17 kDa or virB gene. PCR product of 17 kDa or VirB protein gene which is the well-known antigenic protein for B. henselae was cloned and then express VirB protein. Our VirB specific protein was successfully expressed in pET28a E. coli expression system. Purified VirB protein was then used for development of ELISA for evaluation of antibody to B. henselae from Thai cat blood samples. However, VirB protein in the present study is still need more validation and concerning on the specificity for B. henselae ELISA test for cats due to the cross reactivity among Bartonella spp., different strain of B. henselae, and other bacterial species.