Subtyping Listeria monocytogenes through the combined analyses of genotype and expression of the hlyA virulence determinant.
OBJECTIVE
A major challenge for Listeria monocytogenes diagnostics is that this bacterium is ubiquitous in the environment, and that only a small fraction of the lineages are potential human pathogens. The aim of this work was to obtain a better subtyping of L. monocytogenes through utilization of combined analyses of genotype and the expression of the virulence determinant hlyA.


RESULTS
We investigated the effect of growth temperature and medium on the hlyA expression. The gene expression levels were determined by real-time quantitative reverse transcription PCR. The expression pattern of hlyA was highly diverse among the different strains tested. The expression ranged from repression to a 1000-fold induction for growth at 42 degrees C, as compared with 0 degrees C. The expression patterns were compared with the corresponding genotypes. There were surprisingly low correlations between the expression patterns and the genotype clusterings. This is exemplified for the virulent type strain NTNC 7973 and non-virulent type strain DSMZ 20600. These strains are genetically nearly identical, while the hlyA gene expression patterns are very different.


CONCLUSIONS
The hlyA gene expression was highly diverse even within genetically clustered subgroups of L. monocytogenes. Consequently, the gene expression patterns can be used to further differentiate the strains within these genetic subgroups.


CONCLUSIONS
A major limitation in the control of L. monocytogenes is that the current tools for subtyping are not accurate enough in determining the potential virulent strains. The impact of this study is that we have developed a subtyping approach that actually targets a virulence property.
Rudi
K
K
MATFORSK, Norwegian Food Research Institute, As, Norway. knut.rudi@matforsk.no
Nogva
H K
HK
Naterstad
K
K
Drømtorp
S M
SM
Bredholt
S
S
Holck
A
A
D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism; D015373:Bacterial Typing Techniques / Q000379:methods; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D005838:Genotype; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006460:Hemolysin Proteins; D006801:Humans; D008089:Listeria monocytogenes / Q000145:classification* / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013696:Temperature; D014774:Virulence
D001427:Bacterial Toxins; D003470:Culture Media; D004269:DNA, Bacterial; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; C038705:hlyA protein, Listeria monocytogenes