Single nucleotide polymorphisms and community-associated methicillin-resistant
The present invention is based on the discovery of novel polymorphisms (SNPs) in the penicillin binding protein (pbp3) gene in Staphylococcus aureus . The presence of G88A and/or G2047A SNPs provides an accurate, reliable biomarker for the presence of Methicillin Resistant Staphylococcus aureus (MRSA), specifically the community-associated MRSA (CA-MRSA). The present invention provides reagents used for detecting the SNPs as well as methods of identifying and using these variants to screen subjects for presence of CA-MRSA. The methods involve isolating a biological sample from a mammal (preferably a human) and testing for the presence of a SNP in the pbp3 gene which is associated with CA-MRSA.
1. A method of detecting the presence of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) in a human, comprising the steps of:
a) obtaining a biological sample from a human;
b) isolating genomic material from said biological sample;
c) detecting the presence of a SNP in said genomic material selected from the group consisting of G88A and G2047A present in the penicillin binding protein 3 gene, wherein the detection of said SNP is indicative of the presence of CA-MRSA in said biological sample in said human and,
d) said detecting step is performed by a polymerase chain reaction (PCR), wherein said PCR is selected from one of the following a real-time PCR, a PCR followed by sequencing, a PCR followed by pyrosequencing or an allele-specific PCR, and
e) said PCR is performed using a primer set that comprises SEQ ID 5 and SEQ ID 6.
2. The method of claim 1 , wherein said SNP is G88A.
3. The method of claim 1 , wherein said SNP is G2047A.
4. The method of claim 1 , wherein said biological sample is whole blood, plasma, urine, or exudate from an infected site.
5. The method of claim 1 , wherein said isolating step is performed using guanidinium hydrochloride.