Patent ID: 8518646

Claim:
A method of specifically detecting the presence of a Staphylococcus aureus ( S. aureus ) strain and identifying a methicillin-resistant S. aureus strain from a sample in a single assay, comprising: simultaneously contacting said sample with at least one primer pair that is capable of amplifying a sequence from a nuc gene S. aureus when an S. aureus strain is present in the sample to produce a nuc amplicon, said primer pair comprising a first and a second nuc primer, said first and second nuc primers being between 11 and 45 nucleotides in length and being capable of annealing under PCR conditions to opposite strands of S. aureus chromosomal DNA within the nuc gene; and at least one MREJ primer pair comprising a first and second MREJ primer that is capable of amplifying a polymorphic mec right extremity junction (MREJ) sequence of a methicillin resistant S. aureus (MRSA) strain when an MRSA strain is present in the sample to produce an MREJ amplicon, said MREJ sequence comprising polymorphic sequences from the right extremity of an SCCmec cassette inserted into the S. aureus chromosome and S. aureus chromosomal DNA, said first and second MREJ primers being between 11 and 45 nucleotides in length and being capable of annealing under PCR conditions to opposite strands of MRSA chromosomal DNA, to create an amplification mixture, wherein said PCR conditions comprise 4 mM MgCl 2 , 100 mM Tris (pH 8.3), 10 mM KCl, 5 mM (NH 4 ) 2 SO 4 , 0.15 mg/mL BSA, 4% trehalose at 59° C. subjecting the amplification mixture to an amplification protocol to enable amplification of both nuc and MREJ sequences, thereby producing an amplified sample; and detecting the presence and/or amount of nuc amplicons and MREJ amplicons in the amplified sample as an indication of the presence and or amount of S. aureus and MRSA, respectively, present in the sample.