Compositions for use in identification of bacteria
View Patent ↗The present invention provides compositions, kits and methods for rapid identification and quantification of bacteria by molecular mass and base composition analysis.
1. An oligonucleotide primer pair comprising a forward and a reverse primer, wherein said forward primer comprises a sequence 98% identical to SEQ ID NO: 418, and said reverse primer comprises a sequence 98% identical to SEQ ID NO: 1301.
2. The oligonucleotide primer pair of claim 1 wherein said forward primer is SEQ ID NO: 418.
3. The oligonucleotide primer pair of claim 1 wherein said reverse primer is SEQ ID NO: 1301.
4. The oligonucleotide primer pair of claim 1 wherein at least one of said forward primer and said reverse primer comprises at least one modified nucleobase.
5. The oligonucleotide primer pair of claim 4 wherein at least one of said at least one modified nucleobase is a mass modified nucleobase.
6. The oligonucleotide primer pair of claim 5 wherein said mass modified nucleobase is 5-Iodo-C.
7. The composition of claim 5 wherein said mass modified nucleobase comprises a molecular mass modifying tag.
8. The oligonucleotide primer pair of claim 4 wherein at least one of said at least one modified nucleobase is a universal nucleobase.
9. The oligonucleotide primer pair of claim 8 wherein said universal nucleobase is inosine.
10. The oligonucleotide primer pair of claim 1 wherein at least one of said forward primer and said reverse primer comprises a non-templated T residue at its 5′ end.
11. A kit for identifying, determining one or more characteristics of, or detecting a Staphylococcus aureus bioagent comprising the oligonucleotide primer pair of claim 1 and at least one additional primer pair designed to hybridize to a Staphylococcus aureus gene encoding arcC, aroE, gmk, tpi, yqi, or a combination thereof.
12. The kit of claim 11 further comprising at least one other additional primer pair designed to hybridize to a Staphylococcus aureus gene encoding mecA, mecR1, pvluk, or a combination thereof.
13. The kit of claim 11 wherein said at least one additional primer pair comprises SEQ ID NOs: 437:1232, SEQ ID NOs:590:891, SEQ ID NOs:474:869, SEQ ID NOs: 268:1284, SEQ ID NOs: 318:1300, SEQ ID NOs: 440:1076, SEQ ID NOs: 219:1013, or a combination thereof.
14. The kit of claim 11 wherein said oligonucleotide primer pair of claim 1 and said at least one additional primer pair consists of eight oliconucleotide primer pairs having at least 70% sequence identity with the primer pairs represented by SEQ ID NOs: 437:1232, SEQ ID NOs: 590:891, SEQ ID NOs: 474:869, SEQ ID NOs: 268:1284, SEQ ID NOs: 418:1301, SEQ ID NOs: 318:1300, SEQ ID NOs: 440:1076, and SEQ ID NOs: 219:1013.