Detection of echinocandin-resistant candida glabrata
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Serv
InventorsCau Dinh Pham, Shawn Robert Lockhart
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Abstract
Probes and primers are disclosed for detecting a C. glabrata resistant to an echinocandin in a sample. Method are also disclosed that utilize these probes and primers, wherein the methods can be used to detect a C. glabrata resistant to an echinocandin in a sample.
Claims33
A method for detecting a C. glabrata resistant to an echinocandin in a sample, comprising
- contacting a biological sample comprising DNA with unequal amounts of a forward primer and a reverse primer that can be used to amplify a FKS1-hot spot 1 (HS1) nucleic acid and unequal amounts of a forward and a reverse primer that can be used to amplify a FKS2-HS1 nucleic acid;
- performing asymmetric polymerase chain reaction (PCR) to form amplified DNA product;
- and
- contacting the amplified DNA product with a set of probes, wherein each probe in the set of probes specifically binds FKS1-HS1 DNA mutation or specifically binds a FKS2-HS1 DNA mutation, wherein each of the probes is labeled with a unique label such that the identity of each probe is known by detecting the label;
- detecting amplified DNA product hybridized to the set of probes;
- and
- determining if a probe in the set of probes specific for the FKS1-HS1 DNA mutation or a probe in the set of probe specific for an FKS2-HS1 DNA mutation is bound to the amplified DNA product, wherein binding of the probe specific for the FKS1-HS1 DNA or the probe specific for an FKS2-HS1 DNA mutation indicates that the C. glabrata is resistant to the echinocandin is present in the sample.
An oligonucleotide primer of 20 to 26 nucleotides in length comprising the nucleic acid sequence set forth as one of SEQ ID NOs:
- 15-18, wherein the primer is conjugated to a label.
An oligonucleotide probe of 17 to 30 nucleotides in length comprising the nucleic acid sequence set forth as one of SEQ ID NOs:
- 3-14 or 19-21, wherein the probe is conjugated to a label.
A kit comprising
- a container comprising an oligonucleotide probe of 17 to 30 nucleotides in length, wherein the oligonucleotide probe comprises the nucleic acid sequence set forth as one of SEQ ID NOs:
- 3-14 or 19-21;
- and
- instructions for using the kit.
Dependencies are inferred from the claim text.