Submitted to: Veterinary Microbiology
Publication Type: Review article
Publication Acceptance Date: 9/20/2002
Publication Date: 12/20/2002
Citation: BRICKER, B.J. PCR AS A DIAGNOSTIC TOOL FOR BRUCELLOSIS. VETERINARY MICROBIOLOGY. 2002. v. 90. p. 435-446 Interpretive Summary: This manuscript reviews the historical development of rapid DNA diagnostic tests for brucellosis in animals and humans. The major strategies are described and compared in detail, and their strengths and weaknesses are discussed. The analysis concludes that significant progress has been achieved, but there is a need for further development in the areas of sample preparation, strain differentiation, and reproducibility.
Technical Abstract: Numerous PCR-based assays have been developed for the identification of Brucella to improve diagnostic capabilities. Collectively, the repertoire of assays addresses several aspects of the diagnostic process. For some purposes, the simple identification of Brucella is adequate (e.g. diagnosis of human brucellosis or contamination of food products). In these cases, a genus-specific PCR assay is sufficient. Genus-specific assays tend to be simple, robust, and somewhat permissive of environmental influences. The main genetic targets utilized for these applications are the Brucella BCSP31 gene and the 16S rRNA operon. Other instances require identification of the Brucella species involved. For example, most government-sponsored brucellosis eradication programs include regulations that stipulate a species-specific response. For epidemiological trace-back, strain-specific identification is helpful. Typically, differential PCR-based assays tend to be more complex and consequently more difficult to perform. Several strategies have been explored to differentiate among Brucella species and strains, including locus specific multiplexing (e.g. AMOS-PCR), PCR-RFLP (e.g. the omp2 locus), arbitrary-primed PCR, and ERIC-PCR to name a few. This paper reviews some of the major advancements in molecular diagnostic for Brucella including the development of procedures designed for the direct analysis of a variety of clinical samples. While this progress to date is impressive, there is still room for improvement.