Detection and identification of Salmonella and Shigella
AssigneeUniversity of Hawaii
InventorBruce Roll
Life of this patent
Estimating the term…
Abstract
The present invention is directed to methods and reagents for the specific detection and presumptive identification of various bacteria associated with waterborne infectious disease. In particular, this invention relates to methods and reagents for the specific detection and identification of Salmonella and Shigella in environmental samples such as water and sewage.
Claims14
A method for amplification of nucleic acid from one or more enteric pathogens comprising the steps of:
- a) providing a test sample suspected of containing amplifiable nucleic acid from said one or more enteric pathogens;
- b) isolating said amplifiable nucleic acid from said test sample;
- c) combining said amplifiable nucleic acid with amplification reagents, and at least two primers selected from the group consisting of primers having the nucleic acid sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6 and SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 to form a reaction mixture;
- and
- d) combining said reaction mixture with an amplification enzyme under conditions wherein said amplifiable nucleic acid is amplified to form amplification product.
A method for amplification of nucleic acid from a plurality of enteric pathogens comprising the steps of:
- a) providing a test sample suspected of containing amplifiable nucleic acid of said plurality of enteric pathogens;
- b) isolating said amplifiable nucleic acid from said test sample;
- c) combining said amplifiable nucleic acid with amplification reagents, and all of the primers set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:6 and SEQ ID NO:7, to form a reaction mixture;
- d) combining said reaction mixture with an amplification enzyme under conditions wherein said amplifiable nucleic acid is amplified to form amplification products;
- and
- e) detecting said amplification products.
A method for amplification of nucleic acid from an enteric pathogen comprising the steps of:
- a) providing a test sample suspected of containing amplifiable nucleic acid of said enteric pathogen;
- b) isolating said amplifiable nucleic acid from said test sample;
- c) combining said amplifiable nucleic acid with amplification reagents, and the primers set forth in SEQ ID NO:1 and SEQ ID NO:2, to form a first reaction mixture;
- d) combining said reaction mixture with an amplification enzyme under conditions wherein said amplifiable nucleic acid is amplified to form a first amplification product;
- e) combining said first reaction mixture with amplification reagents, and the primers set forth in SEQ ID NO:3, and SEQ ID NO:4, to form a second reaction mixture;
- f) combining said second reaction mixture with an amplification enzyme under conditions wherein said amplifiable nucleic acid is amplified to form a second amplification product;
- and
- g) detecting said first or second amplification product.
Dependencies are inferred from the claim text.