DETECTION AND SPECIATION OF BACTERIA THROUGH PCR USING UNIVERSAL MAJOR COLD-SHOCK PROTEIN PRIMER OLIGOMERS

The detection of bacteria using PCR is a well-established diagnostic t echnique. However, conventional PCR requires the use of DNA primer oli gomers that are specific to the target organism and, as a consequence, a sample can only be tested for the presence of that specific target. A significant advantage would be to probe a sample for the presence o f any bacteria, followed by identification. To achieve this it is nece ssary to identify a DNA sequence common to all bacteria. Here we demon strate that such a sequence may be that encoding the major cold-shock proteins. Using two universal PCR primer of oligomers from conserved r egions of these gene homologues, we have amplified a 200 base-pair DNA sequence from more than 30 diverse Gram-positive and Gram-negative ba cteria, Including representatives from the genera Aeromonas, Bacillus, Citrobacter, Enterobacter, Enterococcus, Escherichia, Klebsiella, Lac tobacillus, Lactococcus, Listeria, Pediococcus, Photobacterium, Proteu s, Salmonella, Shigella, Staphylococcus, Streptococcus, and Yersinia. Sequence analysis of the amplified products confirmed a high level of DNA homology. Significantly, however, there are sufficient nucleotide variations to allow the unique allocation of each amplified sequence t o its parental bacterium.