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For a long time Salmonella enterica serovar Enteritidis remains the main pathogen of salmonellosis in Russia. Traditionally, different typing methods are used for outbreak investigation to establish the identity of isolates, to identify the way and factors of transmission. Despite of all their advantages, pulse filed gel electrophoresis (PFGE) and whole genome sequencing, which are the “gold typing standards”, are currently unavailable for the majority of the bacteriological laboratories because of their complexity and high requirements for the staff qualification. The aim of this work was to assess the possibility to use the RAPD-PCR for the rapid genotyping of S. Enteritidis strains isolated during outbreaks. 27 strains of S. Enteritidis isolated from patients involved in two unrelated Salmonella outbreaks were studied. 13 strains were isolated during the first outbreak, 14 strains — during the second. For RAPD-PCR four oligonucleotide primers were used: mnv-45, mnv-3-1, mnv-3-2 and RAPD4. Amplified fragments were separated by electrophoresis in a 1.5% agarose gel in 0.5 x TBE-buffer. Analysis of electrophoresis pictures was carried out using Bio-Rad Universal Hood II Gel Doc System. All experiments with Salmonella strains were performed in three repeats to confirm the result reproducibility. Primer mnv-3-2 had the best discriminating ability and allowed to separate the strains belonging to different outbreaks by the number and length of amplified fragments. Primers mnv-3-1, mnv-45 and RAPD4 gave the almost identical profiles of amplified fragments, and mnv-45 also required a special electrophoretic conditions, since the amplified fragments differed slightly (by 20–30 base pairs). The obtained results allow us to recommend the use of RAPD-PCR method with mnv-3-2 primer for operative genotyping of S. Enteritidis isolates, which reveals the individual characteristics of epidemic strains. If the laboratory is equipped with PCR equipment with electrophoretic detection, the strain typing can be performed within one working day.

THE USE OF RAPD-PCR FOR GENOTYPING OF S. ENTERITIDIS ISOLATED DURING THE OUTBREAK