Genotypowanie metodą REA-PFGE pałeczek Listeria monocytogenes izolowanych z próbek materiału klinicznego, próbek środowiskowych i próbek żywności
[Genotyping by REA-PFGE of Listeria monocytogenes isolated from clinical, environmental and food samples]

Streszczenie

Pałeczki Listeria monocytogenes poddano genetycznemu typowaniu metodą REA – PFGE z restryktazami AscI oraz ApaI oraz genotypowaniu metodą multipleks PCR wg Doumitha i wsp. Wśród badanych szczepów wyróżniono 62 profile PFGE w dwóch grupach filogenetycznych. Połączona analiza profili PFGE dla enzymów AscI i ApaI wyodrębniła 8 pulsotypów grupujących dwa lub więcej szczepów pałeczek L. monocytogenes.

Abstract

Listeria monocytogenes strains isolated from clinical food and environmental samples were genotyped by Restriction Enzyme Analysis with Pulsed Field Gel Electrophoresis (REA - PFGE) using ApaI and AscI enzymes according to PulseNet Europe procedure. Analysis of DNA fragments
profiles obtained by AscI digestion demonstrated presence of 62 REA-PFGE profiles grouped in 2 lineages (FI, FII). Diversity of strains source among both lineages was observed. Statistical analysis showed, that strains isolated from clinical samples more frequently are included to lineage FI, then lineage FII. Non-clinical strains were more frequently included to lineage FII. Combined analysis of REA-PFGE profiles for ApaI and AscI enzymes showed 8 unique pulsotypes characteristic for two or more L. monocytogenes isolates.Moreover researched L. monocytogenes strains were analyzed by multiplex-PCR according Doumith et al methodology. PCR-group 4B was most frequent among strains isolated from clinical samples. Correlation between PCR-group and pulsotype was observed only in few cases.