Methods and Compositions for Detecting Virulent and Avirulent Escherichia coli Strains
AssigneeThe Florida State University Research Foundation, Inc.
InventorPrashant Singh
Life of this patent
Estimating the term…
Abstract
Disclosed herein are methods for detecting virulent Shiga toxin-producing E. coli (STEC) strains O26, O103, O121, and O111 in a biological sample comprising the steps of: (i) enriching the bacterial concentration of the biological sample to result in an enriched sample; (ii) isolating DNA from said enriched biological sample; and (iii) detecting virulent strain in said isolated DNA sample via real-time PCR and a melt curve assay. Also disclosed are primers for said assay, as well as kits comprising said primers.
Claims18
1-10. (canceled)
A method for detecting virulent Shiga toxin-producing E. coli (STEC) O111 strain in a biological sample comprising the steps of:
- (i) enriching bacterial concentration of the biological sample to result in an enriched sample;
- (ii) isolating DNA from said enriched biological sample;
- and
- (iii) detecting virulent O111 in said isolated DNA sample;
- wherein detection occurs via real-time PCR high-resolution melt assay or probe-based assay. Wherein said real-time PCR comprises at least one primer pair, wherein said primer pair comprises SEQ ID NO:
- 3 and SEQ ID NO:
- 4, and further wherein assay comprises one or more probes, wherein said probe comprises SEQ ID NO:
- 13 or 14, or both.
20-39. (canceled)
An isolated nucleic acid sequence comprising 90% or more identity to SEQ ID NO:
- 3, 4, 13 or 14.
The isolated nucleic acid sequence of claim 39, wherein the sequence comprises at least 95% identity to SEQ ID NO:
- 3, 4, 13 or 14.
43-45. (canceled)
A kit comprising a primer pair, wherein the primer pair comprises SEQ ID NO:
- 3 and SEQ ID NO:
- 4 and wherein the kit further comprises SEQ ID NO:
- 13 and 14.
48-53. (canceled)
Dependencies are inferred from the claim text.