Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9487836-B2
Publication Date
2016-11-08
Expiration Date
Abstract
The invention relates to a method of identifying a specific yeast species in patient tissue or body fluid. The method comprises the steps of extracting and recovering DNA of the yeast species from the patient tissue or body fluid, amplifying the DNA, hybridizing a probe to the DNA to specifically identify the yeast species, and specifically identifying the yeast species. The invention also relates to a method of identifying a yeast mycotoxin in patient tissue or body fluid. The method comprises the steps of extracting and recovering the yeast mycotoxin from the patient tissue or body fluid, contacting the yeast mycotoxin with an antibody directed against the yeast mycotoxin, and identifying the yeast myocotoxin. Both of these methods can be used to determine if a patient is at risk for or has developed a disease state related to a yeast infection, and to develop an effective treatment regimen for the patient.
Core Innovation
The invention provides a method of identifying a specific yeast species in patient tissue or body fluid by extracting and recovering DNA of the yeast species from the patient tissue or body fluid. The extracted DNA is amplified, and a probe is hybridized to the DNA to specifically identify the yeast species. The probe sequence consists of SEQ ID NO: 7 or a full complement of the sequence to specifically identify Candida kruseii.
The disclosed approach further includes hybridizing a probe to DNA to specifically identify yeast species using probe sequences selected from SEQ ID NO: 1, SEQ ID NO: 4, and SEQ ID NO: 10, including full complementary sequences. The probe format may include fluorescent labeling and is optionally provided using a fluorochrome-dyed bead. Amplification can be implemented as PCR and real-time PCR, and detection can be performed using probe hybridization.
In addition, the invention discloses a method of extracting and recovering yeast mycotoxins from patient samples and detecting them using antibodies directed against the mycotoxin. The detection can be implemented using ELISA, and can optionally use a fluorochrome-coupled bead format. Mycotoxins explicitly include gliotoxin and patulin.
The document describes clinical relevance through validation and example results, including patient urine testing demonstrating detection of Candida albicans and Candida glabrata in asymptomatic patients. It also references clinical assay validation on clinical fluids including urine, CSF, and blood, and provides kit and purified nucleic acid reagent embodiments for performing the nucleic-acid and mycotoxin detection workflows.
Claims Coverage
The partial content identifies one independent claim (clm-00001). The main inventive features cover DNA extraction and recovery from patient tissue/body fluid, DNA amplification, and probe hybridization using a defined SEQ ID NO to specifically identify Candida kruseii; dependent aspects include specified probe sequence options, selectable sample types, and amplification and probe formats.
DNA extraction and recovery from patient tissue or body fluid for yeast species identification
Extracting and recovering DNA of the yeast species from the patient tissue or body fluid.
Amplification of recovered yeast DNA
Amplifying the DNA.
Probe hybridization using SEQ ID NO to specifically identify Candida kruseii
Hybridizing a probe to the DNA to specifically identify the yeast species, wherein the probe sequence consists of SEQ ID NO: 7 or a full complement of the sequence and specifically identifying Candida kruseii.
Probe format using fluorochrome-dyed bead
Hybridizing a probe to the DNA using a probe format wherein the probe is bound to a bead that is dyed with a fluorochrome.
Additional probe sequences for identifying other yeast species
Hybridizing a probe to the DNA to specifically identify other yeast species, using a probe sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 4, and SEQ ID NO: 10, or a full complementary sequence of any of those.
Specified body fluids for the patient sample
Selecting the body fluids from the group consisting of urine, nasal secretions, nasal washes, bronchial lavages, bronchial washes, spinal fluid, sputum, gastric secretions, seminal fluid, other reproductive tract secretions, lymph fluid, whole blood, serum, or plasma.
PCR amplification
Amplifying DNA using PCR.
Real-time PCR amplification
Performing the amplification using real-time PCR.
Overall, the claims coverage centers on extracting and recovering yeast DNA from patient tissue/body fluid, amplifying the DNA, and hybridizing a probe defined by SEQ ID NO: 7 (or its full complement) to specifically identify Candida kruseii, with dependent claim refinements covering other specific SEQ ID probe sequences, enumerated body fluid sample types, and PCR/real-time PCR and fluorochrome-dyed bead probe formats.
Stated Advantages
Supports determining patient risk/disease state for yeast infection.
Supports treatment regimen development.
Documented Applications
Diagnosing yeast infections by identifying specific yeast species (including Candida kruseii and other Candida species) from patient tissue or body fluid.
Detecting yeast mycotoxins from patient samples by antibody-based detection of gliotoxin and patulin.
Clinical testing using patient urine samples, including detection of Candida albicans and Candida glabrata in asymptomatic patients.
Use of assay validation on clinical fluids including urine, CSF, and blood.
Interested in licensing this patent?