Patent ID: 10662484

Abstract:
Provided herein are biological assay mixtures for use in a real-time PCR and High Resolution Melt Analysis (HRMA) comprising: a universal Gram-positive forward and reverse primer pair; a universal Gram-negative forward and reverse primer pair; at least two biological probes, each one comprising a fluorophore, able to detect an amplicon prepared by the universal primer pairs; and an oligonucleotide intercalating dye for the HRMA. Methods for detecting a plurality of bacterial strains in the assay, comprise: contacting a biological sample with the biological assay mixture; preparing a plurality of amplicons for both Gram-positive and Gram-negative bacteria using one of the universal primer pairs; and detecting the amplicons using a plurality of biological probes comprising a fluorophore of one or more color emitters able to hybridize to known bacterial strains, wherein if no bacterial strain is identified, then determining whether Gram-positive and Gram-negative strains exist by using HRMA.