# US 2014/0154694 A1 · Methods, compositions and kits for detection and analysis of antibiotic-resistant bacteria

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- Number: US 2014/0154694 A1
- Document ID: US20140154694A1
- Status: Published application
- Kind: A1
- Application number: 14/149,568
- Filed: 2014-01-07
- Published: 2014-06-05
- Earliest claimed: 2007-04-19
- Assignees: Molecular Detection, Inc.
- Applicants: Molecular Detection, Inc.
- Inventors: Yosef PAITAN
- Web page: https://patentgrep.com/patent/US20140154694A1

## CPC

- C12Q 1/689: for bacteria
- C12Q 2600/16: Primer sets for multiplex assays
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## 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

### Claim 1

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.

### Claim 2 (depends on 1)

A method according to claim 1, wherein said first set of primers comprises at least one of SEQ ID NOs: 10-11.

### Claim 3 (depends on 1)

A method according to claim 1, wherein said second set of primers comprises at least one of SEQ ID NOs: 13-14 and 18-19.

### Claim 4 (depends on 1)

A method according to claim 1, wherein said multi-cycle amplification reaction is a real-time polymerase chain reaction.

### Claim 5 (depends on 1)

A method according to claim 1, wherein said S. aureus-specific polynucleotide sequence is at least a portion of a gene, which is a member selected from nuc and Sa442.

### Claim 6

A kit for identifying MRSA in a sample, said kit comprising:
(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 an S. aureus-specific polynucleotide sequence, wherein said S. aureus-specific polynucleotide sequence is not an orfX polynucleotide;
(c) at least one member selected from: a DNA polymerase enzyme, dNTPs, magnesium, and a stabilizer.