US 12,146,191 B1
Detection and classification of IMP carbapenemases using real-time polymerase chain reaction assay
Nancy D. Hanson, Omaha, NE (US); Hayden Smith, Beaverton, OR (US); and Stacey Morrow, Omaha, NE (US)
Assigned to Creighton University, Omaha, NE (US)
Filed by Creighton University, Omaha, NE (US)
Filed on May 3, 2023, as Appl. No. 18/311,703.
Application 18/311,703 is a division of application No. 16/794,754, filed on Feb. 19, 2020, abandoned.
Claims priority of provisional application 62/807,328, filed on Feb. 19, 2019.
Int. Cl. C12Q 1/6876 (2018.01); C12Q 1/34 (2006.01)
CPC C12Q 1/6876 (2013.01) [C12Q 1/34 (2013.01)] 10 Claims
 
1. A method comprising:
introducing a DNA-containing sample to a first mixture and to a second mixture, the first mixture including a first plurality of primer sets, wherein the first plurality of primer sets includes a first primer comprising the nucleotide sequence of SEQ ID NO:1, a second primer comprising the nucleotide sequence of SEQ ID NO:2, a third primer comprising the nucleotide sequence of SEQ ID NO:3, a fourth primer comprising the nucleotide sequence of SEQ ID NO:4, a fifth primer comprising the nucleotide sequence of SEQ ID NO:5, and a sixth primer comprising the nucleotide sequence of SEQ ID NO:6 first mixture, the second mixture including a second plurality of primer sets differing from the first plurality of primer sets, at least one of the first mixture and the second mixture including a fluorescent dye;
performing a multiplex polymerase chain reaction over multiple cycles with the DNA-containing sample and each of the first mixture and the second mixture; and
detecting fluorescence resulting from the multiplex polymerase chain reaction for each cycle of the multiple cycles.