Development of a multiple primer RAPD assay as a tool for phylogenetic analysis in Listeria spp. strains isolated from milkproduct associated epidemics, sporadic cases of listeriosis and dairy environments

Abstract

A total of 82 Listeria strains comprising four species were examined by amplification with a multiple primer random amplified polymorphic DNA (RAPD) assay. It was the objective of the study to set up a procedure suitable for analysis of the relationships among strains from milkproduct-associated epidemics, strains from sporadic cases of listeriosis and field strains from dairy products and dairy environments. In a preliminary study, 205 primers, each 10 bp long, were screened for suitability as primers and 44 primers showing reliable and reproducible RAPD patterns at a defined reaction condition were selected. The 82 strains were assigned to 54 RAPD groups positioned in 13 major clusters. Strains isolated during milk product-associated epidemics were found to belong to a single cluster I. Human isolates from sporadic cases of listeriosis predominantly were assigned to four separate clusters. It was found that strains of clinical origin were mainly assigned to other clusters than strains of non-clinical origin.