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Identification of marker genes for important Listeria monocytogenes serotypes as a basis for the development of rapid molecular methods
Project
Project code: BfR-BIOS-08-1322-448
Contract period: 01.04.2010
- 31.12.2010
Purpose of research: Applied research
Listeria monocytogenes can cause listeriosis in humans and animals, a severe infections disease with sporadic occurrence in human but with the highest mortality rate amongst foodborne infections. The vast majority of human listeriosis cases (95 %) are associated with the serotypes 1/2a, 1/2b and 4b. Thus, investigations of prevalences of L. monocytogenes in food samples and of consumer?s risk exposure to the pathogen should include both, species and serotype identification. Several microbiological and molecular methods are available for clear species determination. For serotyping, agglutination test with O and H antigene determination according to SEELIGER seems to be the most reliable method. However, also in case of experienced application this method can fail and alternative meth-ods would be more than welcome, particularly in the presence of serological atypical or non-typeable strains. Molecular methods like multiplex PCR have been described for serotyping of Listeria, but up to date they only enable the identification of serogroups but not of serotypes. The object of this project is to identify marker genes (particularly for the serotypes 1/2a, 1/2b and 4b) that will be the basis for the development of fast molecular methods like Multiplex PCR and Real time PCR.We will use cDNA microarrays to generate gene profiles of the serotypes and to identify marker genes (single or in combination) that target the genetic variability of serotypes. These marker genes can either be present in a single serotype or show sequence differ-ences between serotypes (length, inserts, single nucleotide polymorphisms). In another approach we will revise already known genes identified by MLST, MVLST or MLVA etc. for their suitability as marker genes. Serotype specific sequences could then serve to develop primer pairs and gene probes for PCR and Real time PCR.
Section overview
Subjects
- Food microbiology
- Toxicology
