Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1997-09-29
2000-07-18
Myers, Carla J.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 912, 536 2432, 536 2433, 536 231, C12Q 168, C12P 1934, C07H 2104
Patent
active
060905514
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to the field of detection and/or amplification techniques using oligonucleotide probes or primers, and to their application in testing for the presence or in the identification of bacteria of the genus Listeria.
Listeria are Gram + bacteria belonging to the Listeriaceae family which is subdivided into two genera: the genus Listeria and the genus Brochothrix (Collins et al., 1991, International Journal of Systematic Bacteriology, 41, 240-246). The genus Listeria groups together the following bacterial species: Listeria monocytogenes, Listeria innocua, Listeria seeligeri, Listeria welshimeri, Listeria ivanovii, Listeria murrayi and Listeria grayi. Only the Lysteria monocytogenes species exhibits pathogenicity for man. This species is in particular responsible, in man, for severe pathologies such as meningoencephalites, septicemias or abortions. Moreover, since 1981, the Listeria monocytogenes species has been recognized as the agent responsible for several food intoxication epidemics especially in Canada in 1981, in the United States from 1983 to 1985, in Switzerland from 1983 to 1987 and very recently in France in 1992 and 1993. The mortality rate associated with these epidemics is very high, about a third of the cases resulting in abortion or death.
It is therefore important to develop a bacteriological diagnostic test which is sufficiently specific and sensitive to allow rapid and selective detection of the Listeria monocytogenes species, which can be used in particular in the food industry and in medical diagnostics.
A first generation of rapid tests has been proposed, based on the flow cytometry technique (Donnelly C. W., G. J. Baigent, and E. H. Briggs, 1988: Flow cytometry for automated analysis of milk containing Listeria monocytogenes; J. Assoc. Off. Anal. Chem. 71: 655-658) or on the ELISA (enzyme-linked immunosorbent assay) method (Mattingley J. A., B. T. Butman, M. C. Planck, R. J. Durham, and B. J. Robinson, 1988: Rapid monoclonal antibody-based enzyme-linked 4imunosorbent assay for the detection of Listeria in food products; J. Assoc. Off. Anal. Chem. 71: 679-681). However, these techniques allow only the detection of bacteria belonging to the genus Listeria without any discrimination between the species, and in particular the pathogenic species Listeria monocytogenes.
Other techniques of detection by nucleic acid hybridization, using genetic markers, were then developed. It has thus been shown that certain nucleic acid sequences are specific for the sole L. monocytogenes species. These sequences form in particular part of the genes encoding determinants of virulence, such as the region downstream of the hlyA gene (listeriolysin O) (Mengaud J., M. F. Vicente, J. Chenevert, J. Moniz-Pereira, C. Geoffroy, B. Gicquel-Sanzey, F. Baquero, J. C. Perez-Diaz, and P. Cossart, 1988: Expression in Escherichia coli and sequence analysis of the listeriolysin O determinant of Listeria monocytogenes; Infect. Immun. 56: 766-772) or the iap gene (invasion-associated protein, also called p60, Kohler S., M. Leimeister-Wachter, T. Chakraborty, F. Lottspeich, and W. Goebel, 1990: The gene coding for protein p60 of Listeria monocytogenes and its use as a specific probe for Listeria monocytogenes; Infect. Immun. 58: 1943-1950). However, the use of these markers requires a preliminary step of nucleic acid amplification in order to obtain an appropriate sensitivity of the tests, because each of these genes is only present in a single copy.
The present invention overcomes the disadvantages cited above for the detection of the presence of bacteria of the genus Listeria, and more particularly of the species Listeria monocytogenes, using a genetic marker in a process of detection by nucleic acid hybridization combining specificity, sensitivity and speed.
Bacterial ribosomes contain at least three distinct RNA molecules called 5S, 16S and 23S rRNA. Historically, these names were chosen with reference to their speed of sedimentation which is linked to the size of these RNA molecules.
In the proc
REFERENCES:
patent: 5089386 (1992-02-01), Stackebrandt et al.
patent: 5376528 (1994-12-01), King et al.
Baloga et al. Applied and Environmental Microbiology. 57: 2324-2331, Aug. 1991.
Smith et al. Methods in Enzymology. vol. 155, pp. 261-301, 1987.
Wang, Rong-Fu, et al., "Development of a 16S rRNA-Based Oligomer Probe Specific for Listeria monocytogenes", Applied and Environmental Microbology, vol. 57, No. 12, Dec. 1991, pp. 3666-3670.
System, Appl. Microbiol. vol. 15, 487-501 (1992), "Complete 23S Ribosomal RNA Sequences of Gram-positive Bacteria with a Low DNA G+C Content", Wolfgang Lugwig et al.
FEMS Microbiology Letters 96 219-224 (1992), "Studies on the ribosomal RNA operons of Listeria monocytogenes", D.E. Thompson et al.
Mabilat Claude
Sallen Brunehild
Bio Merieux
Myers Carla J.
LandOfFree
Detection of bacteria of genus Listeria using nucleic probe hybr does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Detection of bacteria of genus Listeria using nucleic probe hybr, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Detection of bacteria of genus Listeria using nucleic probe hybr will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2034949