E.i. Dupont De Nemours and Company

Qualicon Diagnostics LLC

Mark Anton Jensen

Stephen Varkey

Daniel R Demarco

Ron Grosz

Neil Alexander Straus

James William Hazel

Anjana Agarwal

Seth L Blumerman

This invention relates to a rapid method for detection and characterization ofbacteria serotype O157:H7 based on the presence of nucleic acid sequences, in particular, to a PCR-based method for detection, and to oligonucleotide molecules and reagents and kits useful therefore. This method is preferably employed to detectO157:H7 in a food or water sample, such as a beef enrichment. The present invention further relates to replication compositions and kits for carrying out the method of the present invention.

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This invention relates to a rapid method for detection and characterization of STEC bacteria based on the presence of nucleic acid sequences, in particular, to a PCR-based method for detection, and to oligonucleotide molecules and reagents and kits useful therefore. This method is preferably employed to detect STEC bacteria in a food or water sample, such as a beef enrichment. The present invention further relates to isolated polynucleotides, replication compositions, kits, and reagent tablets for carrying out the method of the present invention.

Sequences and their use for detection and characterization of STEC bacteria

Disclosed herein are methods for the identification of the species, serotype, and strain of a microorganism. Also disclosed are primers for use in detecting such microorganisms and kits comprising such primers.

Method to amplify nucleic acids to generate fluorescence labeled fragments of conserved and arbitrary products

A method, diagnostic sequences and primers are provided that are useful in identifying the Listeria monocytogenes and Listeria spp. The method involves identifying a RAPD-amplified DNA fragment common to Listeria monocytogenes, then identifying the most conserved regions of that DNA fragment, and the preparing specific primers useful for detecting the presence of a marker within the fragment whereby that set of primers is then useful in the identification of all Listeria monocytogenes. Markers within the same fragment that are specific to the Listeria genus are also identified and are useful for the identification of all Listeria spp.

Genetic markers and methods for the detection of Listeria monocytogenes

A method for the identification of microorganisms is disclosed, comprising first isolating genomic DNA from the suspect microorganism. The isolated DNA has variable sequences interspersed between highly conserved rDNA sequences. The variable sequences are amplified. The amplification reaction is carried out in such manner as to either amplify the variable sequences interspersed between highly conserved rDNA sequences or to amplify arbitrary genomic regions in conjunction with the variable sequences. The resulting amplified DNA fragments are polymorphic with respect to both size and number in a manner which is specific to species, serotype and strain. The distribution of the polymorphic fragments is analyzed and compared to an established database to determine the species, serotype and strain of the suspect microorganism.

Method for the identification of microorganisms by the utilization of

A method, diagnostic sequences and primers are provided that are useful in the identification of the Escherichia coli 0157:H7 serotype. The method first involves the identification of a RAPD-amplified DNA fragment common to 0157:H7 Escherichia coli, the identification of the most conserved regions of that fragment, and the preparation of specific primers useful for detecting the presence of a marker within the fragment whereby that set of primers is then useful in the identification of all 0157:H7 Escherichia coli.

Genetic markers and methods for the detection of escherichia coli

A method is provided for the selection of diagnostic genetic markers fragments and useful in the identification of bacteria at the genus, species or serotype level. The method first involves the identification of a RAPD polymorphic DNA fragment common to a particular microbial group, the identification of the most conserved regions of that fragment, and the preparation of specific primers useful for detecting the presence of a marker within the fragment whereby that set of primers is then useful in the identification of all members of the chosen microbial group. Also provided is a specific diagnostic marker for Salmonella and primers directed thereto.

Selection of diagnostic genetic markers in microorganisms and use of a

Nucleic acid fragments useful in the detection of Salmonella

An improved method for performing a nested polymerase chain reaction (PCR) amplification of a targeted piece of DNA, wherein by controlling the annealing times and concentration of both the outer and the inner set of primers according to the method disclosed, highly specific and efficient amplification of a targeted piece of DNA can be achieved within one reaction vessel without depletion or removal of the outer primers from the reaction mixture vessel.

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West Chester, PA, United States of America

Mark Anton Jensen