Detection of group B streptococcus
First Claim
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1. A method for detecting the presence or absence of GBS 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 pts primers to produce a pts amplification product if a GBS pts nucleic acid molecule is present in said sample, wherein said hybridizing step comprises contacting said sample with a pair of pts probes, wherein the members of said pair of pts probes hybridize within no more than five nucleotides of each other, wherein a first pts probe of said pair of pts probes is labeled with a donor fluorescent moiety and said second pts probe of said pair of pts 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 pts probe and said acceptor fluorescent moiety of said second pts probe, wherein the presence of FRET is indicative of the presence of GBS in said sample, and wherein the absence of FRET is indicative of the absence of GBS in said sample.
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Abstract
The invention provides methods to detect group B streptococcus (GBS) in biological samples using real-time PCR. Primers and probes for the detection of GBS are provided by the invention. Articles of manufacture containing such primers and probes for detecting GBS are further provided by the invention.
10 Citations
36 Claims
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1. A method for detecting the presence or absence of GBS in a biological sample from an individual, said method comprising:
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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 pts primers to produce a pts amplification product if a GBS pts nucleic acid molecule is present in said sample, wherein said hybridizing step comprises contacting said sample with a pair of pts probes, wherein the members of said pair of pts probes hybridize within no more than five nucleotides of each other, wherein a first pts probe of said pair of pts probes is labeled with a donor fluorescent moiety and said second pts probe of said pair of pts 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 pts probe and said acceptor fluorescent moiety of said second pts probe, wherein the presence of FRET is indicative of the presence of GBS in said sample, and wherein the absence of FRET is indicative of the absence of GBS in said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. An article of manufacture, comprising:
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a pair of pts primers;
a pair of pts probes; and
a donor fluorescent moiety and a corresponding acceptor fluorescent moiety. - View Dependent Claims (24, 25, 26, 27)
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28. A method for detecting the presence or absence of GBS in a biological sample from an individual, said method comprising:
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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 pts primers to produce a pts amplification product if a GBS pts nucleic acid molecule is present in said sample, wherein said hybridizing step comprises contacting said sample with a pts probe, wherein the pts 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 pts probe, wherein the presence or absence of fluorescence is indicative of the presence or absence of GBS in said sample. - View Dependent Claims (29, 30, 31, 32, 33)
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34. A method for detecting the presence or absence of GBS in a biological sample from an individual, said method comprising:
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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 pts primers to produce a pts amplification product if a GBS pts nucleic acid molecule is present in said sample, wherein said dye-binding step comprises contacting said pts 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 GBS in said sample, and wherein the absence of binding is indicative of the absence of GBS in said sample. - View Dependent Claims (35, 36)
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Specification