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

  • US 8,512,954 B2
  • Filed: 06/11/2010
  • Issued: 08/20/2013
  • Est. Priority Date: 04/19/2007
  • Status: Expired due to Fees
First Claim
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1. A method of detecting methicillin-resistant S. aureus (MRSA) in a sample, 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 MSSA-orfX polynucleotide sequence, wherein said MSSA-orfX polynucleotide sequence is from a region of the orfX gene surrounding the position at which an insertion cassette would be inserted in methicillin resistant S. aureus but is not inserted in methicillin susceptible S. aureus;

    (c) providing a third set of primers, wherein said third 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;

    (d) combining said first, second and third set of primers with said sample in a reaction mixture;

    (e) performing a multi-cycle amplification reaction with said reaction mixture;

    (f) determining cycle numbers of appearance of each of said mecA, MSSA-orfX and S. aureus-specific polynucleotide sequences,wherein said cycle numbers indicate whether MRSA is present in a sample.

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