Differentiation of Listeria monocytogenes and Listeria innocua by 16S rRNA genes and intraspecies discrimination of Listeria monocytogenes strains by random amplified polymorphic DNA polymorphisms

Abstract

Differences in the 16S rRNA genes (16S rDNA) which can be used to discriminate Listeria monocytogenes from Listeria innocua have been detected. The 16S rDNA were amplified by polymerase chain reaction with a set of oligonucleotide primers which flank a 1.5-kb fragment. Sequence differences were observed in the V2 region of the 16S rDNA both between L. monocytogenes Scott A and L. innocua and between different L. monocytogenes serotypes. Although L. monocytogenes SLCC2371 had the same V2 region sequence as L. innocua, the two species were different within the V9 region at nucleotides 1259 and 1292, in agreement with previous studies (R.-F. Wang, W.-W. Cao, and M. G. Johnson, Appi. Environ. Microbiol. 57:3666-3670, 1991). Intraspecies discrimination ofL. nwnocytogenes strains was achieved by using the patterns generated by random amplified polymorphic DNA primers. Although some distinction can be made within the L. monocytogenes species by their 16S rDNA sequence, a far greater discrimination within species could be made by generating random amplified polymorphic DNA patterns from chromosomal DNA. By using a number of 10-bp primers, unique patterns for each isolate which in all cases examined differentiate between various L. monocytogenes serotypes, even though they may have the same 16S rRNA sequences, could be generated.

Repetitive element sequence-based PCR for species and strain discrimination in the genus Listeria

“…For identification and typing of Listeria strains several methods based on PCR have been developed. Randomly amplified polymorphic DNA (RAPD) has been applied for the typing of Listeria strains (Mazurier and Wernars 1992;Czajka et al 1993;Macgowan et al 1993;Farber and Addison 1994;Balck et al 1995). Repetitive element sequence-based PCR (REP-PCR) is a recently described method for generating DNA fingerprints that can be used to discriminate between bacterial strains.…”

Random amplification of polymorphic bacterial DNA: evaluation of 11 oligonucleotides and application to food contaminated with Listeria monocytogenes

“…Data for Listeria spp. (Mazurier and Wernars 1992;Mazurier et al 1992a, b;Czajka et a!. 1993) and Escherichia coli K1 (Bingen et al 1993) have been published, but none for Yersinia spp.…”

A fast method for monitoring the colonization rate of lactobacilli in a meat model system

“…The technique known as RAPD has been extensively reported for the typing of pathogenic bacterial species (Mazurier et al 1992;Czajca et al 1993;Lawrence et al 1993;Ralph et al 1993;Ross and Dwyer 1993). In Gram-positive bacteria, it has been used for the typing of staphylococci (Wells and McClelland 1990), Lactococcus lactis strains (Cancilla et al 1992) and lactobacilli (Delley et al 1990;Cocconcelli et al 1995).…”