[Genome analysis and primer sets determination for listeria species detection and gene typing]

Computer analysis of listeria genome isolates of sequences from EMBL, GenBank, DDBJ data bases has been made. Variable and highly conservative (homology degree is 90-100% for all known isolates) genes loci iap and listeriolysin (cytolysin) gene locuses have been determined. Primer sets for detection and differentiation of Listeria species by polymerase chain reaction (PCR) were designed by computer and following thermodynamic analysis. Primer sets can provide detection of Listeria monocytogenes, L. ivanovii, L. seeligeri, L. grayi, L. innocua, L. welshimeri and detection of only pathogenic L. monocytogenes and Listeria species with listeriolysin (cytolysin) gene. Differentiation of 6 Listeria species can be done by means of two primer sets for iap and listeriolysin genes. Amplified fragments of listeria species DNA have different amplicon size. Primers melting temperatures selection allows one to carry out listeria species typing by multiplex PCR in a single tube.