# US 9,434,986 B2 · Methods and kits used in the detection of fungus

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- Number: US 9,434,986 B2
- Document ID: US9434986B2
- Status: Granted
- Kind: B2
- Application number: 14/274,974
- Filed: 2014-05-12
- Published: 2014-09-04
- Granted: 2016-09-06
- Earliest claimed: 2010-01-26
- Assignees: The Translational Genomics Research Institute; Arizona Board of Regents On Behalf of Northern Arizona University
- Applicants: The Translational Genomics Research Institute; Arizona Board of Regents On Behalf of Northern Arizona University
- Inventors: Cindy Liu; Sergey Kachur; Lance Price; Paul Keim
- Web page: https://patentgrep.com/patent/US9434986B2

## CPC

- C12Q 1/6844: Nucleic acid amplification reactions
- C12Q 1/6895: for plants, fungi or algae
- C12Q 2304/00: Chemical means of detecting microorganisms
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The invention encompasses method of using quantitative PCR to detect fungal organisms in clinical and environmental samples to generate standards that allow the quantification of fungal organisms in the samples.

## Claims

### Claim 1

A method of detecting a fungus in a sample, the method comprising the steps of:
adding a first oligonucleotide mixture to an amplification mixture, wherein the first oligonucleotide mixture comprises oligonucleotides comprising sequences of SEQ ID NO. 6 and SEQ ID NO. 7 and the amplification mixture comprises a nucleic acid isolated from the sample;
adding a second oligonucleotide mixture to the amplification mixture, wherein the second oligonucleotide mixture comprises at least two oligonucleotides selected from the group consisting of oligonucleotides comprising sequences of SEQ ID NO. 8, SEQ ID NO. 9, SEQ ID NO. 10, SEQ ID NO. 11, SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 14, and SEQ ID NO. 15;
subjecting the mixture to conditions that allow nucleic acid amplification; and
detecting the presence of the fungus based on the results of the nucleic acid amplification.

### Claim 2 (depends on 1)

The method of claim 1 further comprising adding an oligonucleotide probe to the mixture.

### Claim 3 (depends on 2)

The method of claim 2, wherein the oligonucleotide probe comprises the sequence of SEQ ID NO. 2.

### Claim 4 (depends on 2)

The method of claim 2, wherein the second oligonucleotide mixture comprises oligonucleotides comprising sequences of SEQ ID NO. 8, SEQ ID NO. 9, SEQ ID NO. 10, SEQ ID NO. 11, SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 14, and SEQ ID NO. 15.

### Claim 5 (depends on 1)

The method of claim 1, wherein the sample is from a subject.

### Claim 6 (depends on 5)

The method of claim 5, wherein the subject is a human.

### Claim 7

A method of detecting a fungus in a sample, the method comprising the steps of:
adding a first oligonucleotide mixture that comprises oligonucleotides having sequences of SEQ ID NO. 6 and SEQ ID NO. 7 to an amplification mixture comprising a nucleic acid isolated from the sample;
adding a second oligonucleotide mixture to the amplification mixture, wherein the oligonucleotide mixture comprises oligonucleotides having sequences of SEQ ID NO. 8, SEQ ID NO. 9, SEQ ID NO. 10, SEQ ID NO. 11, SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 14, and SEQ ID NO. 15;
subjecting the mixture to conditions that allow nucleic acid amplification; and
detecting the presence of the fungus.

### Claim 8 (depends on 7)

The method of claim 7, wherein the sample is from a subject.

### Claim 9 (depends on 5)

The method of claim 5, wherein the subject is an animal.

### Claim 10 (depends on 1)

The method of claim 1, wherein the sample is from an environmental source.

### Claim 11 (depends on 8)

The method of claim 8, wherein the subject is a human.

### Claim 12 (depends on 8)

The method of claim 8, wherein the subject is an animal.

### Claim 13 (depends on 7)

The method of claim 7, wherein the sample is from an environmental source.

### Claim 14 (depends on 7)

The method of claim 7 further comprising adding an oligonucleotide probe to the mixture.

### Claim 15 (depends on 14)

The method of claim 14, wherein the oligonucleotide probe comprises the sequence of SEQ ID NO. 2.