Methods, compositions and kits for detection and analysis of antibiotic-resistant bacteria

A method of detecting methicillin-resistant S. aureus (MRSA) in a sample, wherein said sample contains one or a mixture of bacterial species, said method comprising:

• (a) providing a first set of primers, wherein said first set of primers are complementary to at least a portion of a mecA polynucleotide sequence;
• (b) providing a second set of primers, wherein said second set of primers are complementary to at least a portion of an S. aureus-specific polynucleotide sequence, and wherein said S. aureus-specific polynucleotide sequence is not an orfX polynucleotide;
• (c) combining said first, second set of primers with said sample in a reaction mixture;
• (d) performing a multi-cycle amplification reaction with said reaction mixture;
• (e) determining cycle numbers of appearance of each of said mecA and S. aureus-specific polynucleotide sequences,
• (f) comparing said cycle numbers of appearance of said mecA and S. aureus-specific polynucleotide sequences to each other,
• wherein if said mecA polynucleotide sequence has a cycle number of appearance that is substantially the same as that of said S. aureus-specific polynucleotide sequence, then MRSA is present in said sample.