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

• US 8,557,974 B2
• Filed: 05/08/2012
• Issued: 10/15/2013
• Est. Priority Date: 04/19/2007
• Status: Expired due to Fees

• Current Assignee: Molecular Detection Inc.
• Original Assignee: Molecular Detection Inc.
• Inventors: Paitan, Yosef
• Primary Examiner(s): WOOLWINE, SAMUEL C
• Application Number:
  US13/466,997
• Publication Number:
  US 20120225429A1
• Time in Patent Office: 525 Days
• Field of Search: None
• US Class Current: 536/24.33
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

The present invention relates generally to detection of antibiotic-resistant bacteria in a sample. In particular, the invention provides methods, compositions and kits for detecting and analyzing methicillin-resistant Staphylococcus aureus (MRSA) and other methicillin-resistant bacteria in a sample.

Claims

1. A kit that comprises:

(a) a first set of primers complementary to at least a portion of a mecA polynucleotide sequence;

(b) a second set of primers complementary to at least a portion of a bridging region;

(c) a third set of primers complementary to at least a portion of an MSSA-orfX polynucleotide sequence; and

(d) at least one member selected from;

a DNA polymerase enzyme, (dNTPs, magnesium and a stabilizer.

2. A method of identifying bacteria in a sample, said method comprising:

(a) providing a kit according to claim 1;

(b) determining whether a mecA polynucleotide is present in said sample using said first set of primers;

(c) determining whether a bridging region sequence is present in said sample using said second set of primers; and

(d) determining whether an MSSA-orfX polynucleotide is present in said sample using said third set of primers,

wherein the combination of (b), (c) and (d) present in said sample identifies bacteria in said sample.

3. A kit for detecting an antibiotic-resistant bacterial strain in a sample, wherein said kit comprises:

(a) a first set of primers, wherein said first set of primers are capable of producing a first amplification product from at least a portion of a gene, wherein said gene confers antibiotic-resistance;

(b) a second set of primers, wherein said second set of primers are capable of producing a second amplification product from at least a portion of a bridging region;

(c) a third set of primers, wherein said third set of primers are capable of producing a third amplification product from at least a portion of a bacterial strain-specific polynucleotide sequence.