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US 12,680,140 B2Granted
Detection of methicillin-resistant Staphylococcus aureus in biological samples
AssigneeQuest Diagnostics Investments Incorporated
InventorsLakshmi Nair, Heather Vincent, Huong Mai, Michelle Tabb, Maurice Exner
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Abstract
Disclosed are methods of identifying a methicillin-resistant Staphylococcus aureus (MRSA) in a sample wherein the methods involve detecting a S. aureus-specific nucleic acid sequence, mecA and mecC, in the sample. Kits for determining the presence of MRSA in a sample are also provided.
Claims15
A method for determining the presence or absence of methicillin-resistant Staphylococcus aureus (MRSA) in a biological sample comprising:
- (a) contacting the biological sample with:
- (i) a first primer pair that specifically hybridizes under stringent conditions to a segment of a target nucleic acid specific for Staphylococcus aureus, if present, wherein at least one primer of the first primer pair is associated with a fluorophore label,
- (ii) a second primer pair that specifically hybridizes under stringent conditions to a segment of a target mecC nucleic acid, if present, wherein one primer of the second primer pair is a primer-probe that (i) is associated with a fluorophore label and (ii) comprises a probe sequence element of SEQ ID NO:
- 5 at the 5′ end of the primer-probe and SEQ ID NO:
- 6 at the 3′ end of the primer-probe, wherein the probe sequence element further comprises a quencher, and
- (iii) a third primer pair that specifically hybridizes under stringent conditions to a segment of a target mecA nucleic acid, if present, wherein at least one primer of the third primer pair is associated with a fluorophore label, to produce a reaction-sample mixture,
- (b) subjecting the reaction-sample mixture to real-time polymerase chain reaction (PCR) conditions under which each of the target nucleic acids present in the biological sample is amplified to produce a fluorescent signal,
- (c) measuring the amount of fluorescent signal produced by each fluorophore label, and
- (d) determining the presence or absence of MRSA by comparing the amount of fluorescence from the S. aureus-specific target nucleic acid and the mecA and mecC target nucleic acids.
6 more elements
A detection method comprising:
- (a) contacting a biological sample with:
- (i) a first primer pair that specifically hybridizes under stringent conditions to a segment of a target nucleic acid specific for Staphylococcus aureus, if present, wherein at least one primer of the first primer pair is associated with a fluorophore label,
- (ii) a second primer pair that specifically hybridizes under stringent conditions to a segment of a target mecC nucleic acid, if present, wherein one primer of the second primer pair is a primer-probe that (i) is associated with a fluorophore label and (ii) comprises a probe sequence element of SEQ ID NO:
- 5 at the 5′ end of the primer-probe and SEQ ID NO:
- 6 at the 3′ end of the primer-probe, wherein the probe sequence element further comprises a quencher, and
- (iii) a third primer pair that specifically hybridizes under stringent conditions to a segment of a target mecA nucleic acid, if present, wherein at least one primer of the third primer pair is associated with a fluorophore label, to produce a reaction-sample mixture,
- (b) subjecting the reaction-sample mixture to real-time polymerase chain reaction (PCR) conditions under which each of the target nucleic acids present in the biological sample is amplified to produce a fluorescent signal, and
- (c) measuring the amount of fluorescent signal produced by each fluorophore label.
5 more elements
Dependencies are inferred from the claim text.