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US 2014/0274770 A1Published application
Pcr assays and reagents for molecular detection of infectious agents
AssigneeQuidel Corporation
InventorTodd Denison Pack
Life of this patent
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Abstract
The present disclosure is directed to PCR-based assays and compositions for use in molecular detection of viral, bacterial and parasitic infectious agents in body fluid or tissue samples, and in particular to multiplex assays, as well as to solid reagent compositions for use in such assays.
Claims14
A method for identifying the presence or absence of a Clostridium species in a sample, the method comprising:
- a) spiking a sample suspected of containing a target nucleic acid sequence of a Clostridium species with a process control sequence, to form a spiked solution;
- b) exposing the spiked solution to lysing conditions to form a lysed solution;
- c) contacting said lysed solution with an amplification solution to form a mixture, the amplification solution comprising
- (i) at least two PCR analyte primer pairs, each pair specific for a different target nucleic acid sequence of a Clostridium species;
- (ii) a PCR control primer pair specific for said process control sequence;
- (iii) at least one analyte probe specific for each said different target nucleic acid sequence;
- (iv) a control probe specific for said process control sequence;
- (v) a thermostable enzyme having DNA polymerase activity;
- and
- (vi) deoxyribonucleotides dATP, dCTP, dGTP and dTTP or dUTP;
- d) producing an amplicon from at least one target nucleic acid sequence in the mixture, if present, using a single set of thermocycling conditions in a thermocycler;
- and
- e) monitoring the at least one analyte probe to determine the presence or absence of the target nucleic acid sequence.
10 more elements
A composition, comprising:
- at least two PCR analyte primer pairs, each pair substantially complementary to a different target DNA sequence derived from Clostridium difficile;
- a PCR control primer pair substantially complementary to a process control DNA sequence;
- at least one analyte probe for specific binding to each said target DNA sequence;
- a control probe for binding to the process control DNA sequence;
- a thermostable enzyme having DNA polymerase activity;
- and
- deoxyribonucleotides dATP, dCTP, dGTP and dTTP or dUTP;
- wherein said composition is in solid form,
- and wherein said analyte primer pairs and control primer pair are designed such that amplification and detection of said different target DNA sequences and said process control DNA sequence can be performed simultaneously using the same thermal cycling conditions on a thermocycler.
6 more elements
Dependencies are inferred from the claim text.