Methods and compositions for detecting fungi and mycotoxins

• US 10,036,748 B2
• Filed: 06/28/2017
• Issued: 07/31/2018
• Est. Priority Date: 04/01/2006
• Status: Active Grant

• Current Assignee: RTL Purchaser LLC
• Original Assignee: AdvaTect Diagnostics, LLC
• Inventors: Hooper, Dennis G.
• Primary Examiner(s): Hines, Jana A
• Application Number:
  US15/635,807
• Publication Number:
  US 20170336410A1
• Time in Patent Office: 398 Days
• Field of Search: 436 8, 436 15, 436164, 436165, 436166, 436169, 436172, 436518, 436523, 436546, 436800, 435 4, 435 7, 435 71
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/142 Toxicological screening, e.g. expression profiles which identify toxicity
• CPC Class Codes: G01N 2469/10 Detection of antigens from microorganism in sample from host
• CPC Class Codes: G01N 2800/26 Infectious diseases, e.g. generalised sepsis
• CPC Class Codes: G01N 2800/50 Determining the risk of developing a disease
• CPC Class Codes: G01N 33/56961 Plant cells or fungi

Abstract

The invention relates to a method of identifying a specific fungal species in patient tissue or body fluid. The method comprises the steps of extracting and recovering DNA of the fungal species from the patient tissue or body fluid, amplifying the DNA, hybridizing a probe to the DNA to specifically identify the fungal species, and specifically identifying the fungal species. The invention also relates to a method of identifying a mycotoxin in patient tissue or body fluid. The method comprises the steps of extracting and recovering the mycotoxin from the patient tissue or body fluid, contacting the mycotoxin with an antibody directed against the mycotoxin, and identifying the 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 fungal infection, and to develop an effective treatment regimen for the patient.

Claims

1. A method of quantifying a mycotoxin in a patient tissue or a body fluid, the method comprising:

extracting and recovering the mycotoxin from the patient tissue or the body fluid, wherein the mycotoxin is a tricothecene mycotoxin, wherein the body fluid is selected from the group consisting of a urine sample, a nasal secretion sample, a nasal wash sample, and a sputum sample, and wherein the patient has been exposed to mold; and

quantifying the mycotoxin to determine whether the patient has a fungal infection that produces the mycotoxin, wherein the method detects 0.2 ng/ml of the tricothecene mycotoxin.

2. The method of claim 1 further comprising the step of developing an effective treatment regimen for the patient.

3. The method of claim 1 wherein the tissue is derived from a patient tissue biopsy and is in a 10% formalin solution or is in a paraffin block.

4. The method of claim 1 further comprising the step of identifying the mycotoxin using negative and positive control samples.

5. The method of claim 1 further comprising the step of using calibration reagents to quantify the mycotoxin.

6. The method of claim 1 wherein there is no detection of aflatoxins or ochratoxins.

7. A method of determining if a patient is at risk for or has developed a disease state related to a fungal infection that produces a mycotoxin, the method comprising:

extracting and recovering the mycotoxin from a tissue or a body fluid of the patient, wherein the mycotoxin is a tricothecene mycotoxin, wherein the body fluid is selected from the group consisting of a urine sample, a nasal secretion sample, a nasal wash sample, and a sputum sample, and wherein the patient has been exposed to mold; and

quantifying the mycotoxin to determine if the patient is at risk for or has developed the disease stale related to the fungal infection that produces the mycotoxin, wherein the method detects 0.2 ng/ml of the tricothecene mycotoxin.

8. The method of claim 7 further comprising the step of developing an effective treatment regimen for the patient.

9. The method of claim 7 wherein the tissue is derived from a patient tissue biopsy and is in a 10% formalin solution or is in a paraffin block.

10. The method of claim 7 further comprising the step of identifying the mycotoxin using negative and positive control samples.

11. The method of claim 7 further comprising the step of using calibration reagents to quantify the mycotoxin.

12. The method of claim 7 wherein there is no detection of aflatoxins or ochratoxins.