Detection of mecA-containing Staphylococcus spp.
AssigneeMayo Foundation for Medical Education and Research
InventorsJames R. Uhl, Franklin R. Cockerill III
Life of this patent
Estimating the term…
Abstract
The invention provides methods to detect mecA-containing Staphylococcus spp. in biological samples using real-time PCR. Primers and probes for the detection of S. aureus are provided by the invention. Articles of manufacture containing such primers and probes for detecting mecA-containing Staphylococcus spp. are further provided by the invention.
Claims36
A method for detecting the presence or absence of mecA-containing Staphyloccus spp. in a biological sample from an individual, said method comprising:
- performing at least one cycling step, wherein a cycling step comprises an amplifying step and a hybridizing step, wherein said amplifying step comprises contacting said sample with a pair of mecA primers to produce a mecA amplification product if a mecA nucleic acid molecule is present in said sample, wherein said hybridizing step comprises contacting said sample with a pair of mecA probes, wherein the members of said pair of mecA probes hybridize within no more than five nucleotides of each other, wherein a first mecA probe of said pair of mecA probes is labeled with a donor fluorescent moiety and said second mecA probe of said pair of mecA probes is labeled with a corresponding acceptor fluorescent moiety;
- and
- detecting the presence or absence of fluorescence resonance energy transfer (FRET) between said donor fluorescent moiety of said first mecA probe and said acceptor fluorescent moiety of said second mecA probe,
- wherein the presence of FRET is indicative of the presence of one or more mecA-containing Staphylococcus spp. in said sample, and wherein the absence of FRET is indicative of the absence of a mecA-containing Staphylococcus spp. in said sample.
An article of manufacture, comprising:
- a pair of mecA primers;
- a pair of mecA probes;
- and
- a donor fluorescent moiety and a corresponding acceptor fluorescent moiety.
A method for detecting the presence or absence of mecA-containing Staphylococcus spp. in a biological sample from an individual or in a non-biological sample, said method comprising:
- performing at least one cycling step, wherein a cycling step comprises an amplifying step and a hybridizing step, wherein said amplifying step comprises contacting said sample with a pair of mecA primers to produce a mecA amplification product if a mecA nucleic acid molecule is present in said sample, wherein said hybridizing step comprises contacting said sample with a mecA probe, wherein the mecA probe is labeled with a donor fluorescent moiety and a corresponding acceptor fluorescent moiety;
- and
- detecting the presence or absence of fluorescence resonance energy transfer (FRET) between said donor fluorescent moiety and said acceptor fluorescent moiety of said mecA probe,
- wherein the presence or absence of fluorescence is indicative of the presence or absence of mecA-containing Staphylococcus spp. in said sample.
A method for detecting the presence or absence of mecA -containing Staphylococcus spp. in a biological sample from an individual or in a non-biological sample, said method comprising:
- performing at least one cycling step, wherein a cycling step comprises an amplifying step and a dye-binding step, wherein said amplifying step comprises contacting said sample with a pair of mecA primers to produce a mecA amplification product if a mecA nucleic acid molecule is present in said sample, wherein said dye-binding step comprises contacting said mecA amplification product with a double-stranded DNA binding dye;
- and
- detecting the presence or absence of binding of said double-stranded DNA binding dye into said amplification product,
- wherein the presence of binding is indicative of the presence of one or more mecA-containing Staphylococcus spp. in said sample, and wherein the absence of binding is indicative of the absence of a mecA-containing Staphylococcus spp. in said sample.
Dependencies are inferred from the claim text.