Method and kit for identifying antibiotic-resistant microorganisms
First Claim
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1. A device for detecting nucleic acids encoding resistance to an antibiotic, comprising:
- a solid support and a plurality of detectably labeled solution-phase hybridization probes distributed among a plurality of loci thereon, said plurality of loci comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of bacteria in the genus Staphylococcus, comprising Staphylococcus aureus and Staphylococcus epidermidis, but do not hybridize to ribosomal nucleic acids of bacteria in the genus Enterococcus, a second locus that comprises (a) one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of bacteria in the genus Enterococcus, comprising Enterococcus faecalis and Enterococcus faecium, but not Staphylococcus aureus or any other bacteria in the genus Staphylococcus, and (b) at least one probe that hybridizes to ribosomal nucleic acids from Staphylococcus aureus but not from other species in the genus Staphylococcus or bacteria in the genus Enterococcus, a third locus that comprises a probe mix containing a first mecA probe that comprises SEQ ID NO;
2 or the complement thereof, a second mecA probe that comprises SEQ ID NO;
3 or the complement thereof, and a third mecA probe that comprises SEQ ID NO;
5 or the complement thereof, and a fourth locus that comprises a probe mix containing probes that collectively hybridize to VanA nucleic acids and to VanB nucleic acids, wherein said probe mix of said fourth locus has nucleotide sequences that consists of the nucleotide sequences SEQ ID NO;
27 or the complement thereof, SEO ID NO ;
28 or the complement thereof, SEQ ID NO;
29 or the complement thereof, SEQ ID NO;
30 or the complement thereof, SEQ ID NO;
40 or the complement thereof, and SEQ ID NO;
41 or the complement thereof.
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Abstract
The invention provides a rapid sample-processing method for preparing hybridization reaction mixtures substantially depleted of RNA, and a method of identifying the methicillin-resistance status and vancomycin-resistance status of an organism.
18 Citations
22 Claims
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1. A device for detecting nucleic acids encoding resistance to an antibiotic, comprising:
- a solid support and a plurality of detectably labeled solution-phase hybridization probes distributed among a plurality of loci thereon, said plurality of loci comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of bacteria in the genus Staphylococcus, comprising Staphylococcus aureus and Staphylococcus epidermidis, but do not hybridize to ribosomal nucleic acids of bacteria in the genus Enterococcus, a second locus that comprises (a) one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of bacteria in the genus Enterococcus, comprising Enterococcus faecalis and Enterococcus faecium, but not Staphylococcus aureus or any other bacteria in the genus Staphylococcus, and (b) at least one probe that hybridizes to ribosomal nucleic acids from Staphylococcus aureus but not from other species in the genus Staphylococcus or bacteria in the genus Enterococcus, a third locus that comprises a probe mix containing a first mecA probe that comprises SEQ ID NO;
2 or the complement thereof, a second mecA probe that comprises SEQ ID NO;
3 or the complement thereof, and a third mecA probe that comprises SEQ ID NO;
5 or the complement thereof, and a fourth locus that comprises a probe mix containing probes that collectively hybridize to VanA nucleic acids and to VanB nucleic acids, wherein said probe mix of said fourth locus has nucleotide sequences that consists of the nucleotide sequences SEQ ID NO;
27 or the complement thereof, SEO ID NO ;
28 or the complement thereof, SEQ ID NO;
29 or the complement thereof, SEQ ID NO;
30 or the complement thereof, SEQ ID NO;
40 or the complement thereof, and SEQ ID NO;
41 or the complement thereof. - View Dependent Claims (2, 3, 4, 9, 10, 13, 19)
- a solid support and a plurality of detectably labeled solution-phase hybridization probes distributed among a plurality of loci thereon, said plurality of loci comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of bacteria in the genus Staphylococcus, comprising Staphylococcus aureus and Staphylococcus epidermidis, but do not hybridize to ribosomal nucleic acids of bacteria in the genus Enterococcus, a second locus that comprises (a) one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of bacteria in the genus Enterococcus, comprising Enterococcus faecalis and Enterococcus faecium, but not Staphylococcus aureus or any other bacteria in the genus Staphylococcus, and (b) at least one probe that hybridizes to ribosomal nucleic acids from Staphylococcus aureus but not from other species in the genus Staphylococcus or bacteria in the genus Enterococcus, a third locus that comprises a probe mix containing a first mecA probe that comprises SEQ ID NO;
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5. A device for detecting nucleic acids encoding resistance to an antibiotic, comprising:
- a solid support and a plurality of detectably labeled solution-phase hybridization probes distributed among at least five loci thereon, comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of bacteria in the genus Staphylococcus, comprising Staphylococcus aureus, Staphylococcus intermedius and Staphylococcus epidermidis, a second locus that comprises a probe that hybridizes to ribosomal nucleic acids from Staphylococcus aureus but not other species in the genus Staphylococcus, a third locus that comprises a probe mix comprising SEQ ID NO;
2 or the complement thereof, SEQ ID NO;
3 or the complement thereof, and SEQ ID NO;
5 or the complement thereof, a fourth locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of Gram-positive bacteria comprising the high(G+C) subset thereof, a plurality of species of bacteria in the family Enterobacteriaceae, a plurality of species of bacteria in the genus Enterococcus, and a plurality of species of bacteria in the genus Staphylococcus; and
a fifth locus that comprises a probe mix containing probes that collectively hybridize to VanA nucleic acids and to VanB nucleic acids, wherein said probe mix of said fifth locus has nucleotide sequences that consists of the nucleotide sequences SEQ ID NO;
27 or the complement thereof, SEQ ID NO;
28 or the complement thereof, SEQ ID NO;
29 or the complement thereof, SEQ ID NO;
30 or the complement thereof, SEQ ID NO;
40 or the complement thereof, and SEQ ID NO;
41 or the complement thereof. - View Dependent Claims (6, 15, 16, 20)
- a solid support and a plurality of detectably labeled solution-phase hybridization probes distributed among at least five loci thereon, comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of bacteria in the genus Staphylococcus, comprising Staphylococcus aureus, Staphylococcus intermedius and Staphylococcus epidermidis, a second locus that comprises a probe that hybridizes to ribosomal nucleic acids from Staphylococcus aureus but not other species in the genus Staphylococcus, a third locus that comprises a probe mix comprising SEQ ID NO;
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7. A device for detecting nucleic acids encoding resistance to an antibiotic, comprising:
- a solid support and a plurality of detectably labeled solution-phase hybridization probes distributed among a plurality of loci thereon, said plurality of loci comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of bacteria in the genus Enterococcus, comprising Enterococcus faecalis and Enterococcus faecium, a second locus that comprises a probe mix containing probes that collectively hybridize to VanA nucleic acids and to VanB nucleic acids, wherein said probe mix of said second locus has nucleotide sequences that consists of the nucleotide sequences SEQ ID NO;
27 or the complement thereof, SEQ ID NO;
28 or the complement thereof, SEQ 1D NO;
29 or the complement thereof, SEQ ID NO;
30 or the complement thereof, SEQ ID NO;
40 or the complement thereof, and SEQ ID NO;
41 or the complement thereof, and a third locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of Gram-positive bacteria comprising the high(G+C) subset thereof, a plurality of species of bacteria in the family Enterobacteriaceae, a plurality of species of bacteria in the genus Enteroeoccus, and a plurality of species of bacteria in the genus Staphylococcus. - View Dependent Claims (8, 14, 21)
- a solid support and a plurality of detectably labeled solution-phase hybridization probes distributed among a plurality of loci thereon, said plurality of loci comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of bacteria in the genus Enterococcus, comprising Enterococcus faecalis and Enterococcus faecium, a second locus that comprises a probe mix containing probes that collectively hybridize to VanA nucleic acids and to VanB nucleic acids, wherein said probe mix of said second locus has nucleotide sequences that consists of the nucleotide sequences SEQ ID NO;
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11. A device for detecting nucleic acids encoding resistance to an antibiotic, comprising:
- a solid support and a plurality of immobilized detectably labeled probes distributed among a plurality of loci thereon, said plurality of loci comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of bacteria in the genus Staphylococcus, comprising Staphylococcus aureus and Staphylococcus epidermidis, but do not hybridize to ribosomal nucleic acids of bacteria in the genus Enterococeus, a second locus that comprises (a) one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of bacteria in the genus Enterococcus, comprising Enterococcus faecalis and Enterococcus faecium, but not Staphylococcus aureus or any other bacteria in the genus Staphylococcus, and (b) at least one probe that hybridizes to ribosomal nucleic acids from Staphylococcus aureus but not from other species in the genus Staphylococcus or bacteria in the genus Enterococcus, a third locus that comprises a probe mix wherein said probe mix comprises a first mecA probe that comprises SEQ ID NO;
2 or the complement thereof, a second mecA probe that comprises SEQ ID NO;
3 or the complement thereof, and a third mecA probe that comprises SEQ ID NO;
5 or the complement thereof, and a fourth locus that comprises a probe mix containing probes that collectively hybridize to VanA nucleic acids and to VanB nucleic acids, wherein said probe mix of said fourth locus has nucleotide sequences that consists of the nucleotide sequences SEQ ID NO;
27 or the complement thereof, SEQ ID NO;
28 or the complement thereof, SEQ ID NO;
29 or the complement thereof, SEQ ID NO;
30 or the complement thereof, SEQ ID NO;
40 or the complement thereof, and SEQ ID NO;
41 or the complement thereof. - View Dependent Claims (12, 17, 18, 22)
- a solid support and a plurality of immobilized detectably labeled probes distributed among a plurality of loci thereon, said plurality of loci comprising, a first locus that comprises one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of species of bacteria in the genus Staphylococcus, comprising Staphylococcus aureus and Staphylococcus epidermidis, but do not hybridize to ribosomal nucleic acids of bacteria in the genus Enterococeus, a second locus that comprises (a) one or more probes that collectively hybridize to ribosomal nucleic acids from a plurality of bacteria in the genus Enterococcus, comprising Enterococcus faecalis and Enterococcus faecium, but not Staphylococcus aureus or any other bacteria in the genus Staphylococcus, and (b) at least one probe that hybridizes to ribosomal nucleic acids from Staphylococcus aureus but not from other species in the genus Staphylococcus or bacteria in the genus Enterococcus, a third locus that comprises a probe mix wherein said probe mix comprises a first mecA probe that comprises SEQ ID NO;
Specification