Methods and compositions for detecting fungi and mycotoxins
View Patent ↗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.
1. A method of quantifying a macrocyclic tricothecene mycotoxin in a patient tissue or a body fluid, the method comprising the steps of: extracting a recovering the tricothecene mycotoxin from the patient tissue or the body fluid wherein the body fluid is selected from the group consisting of a urine sample, a nasal secretion sample, a nasal wash sample, and sputum sample and wherein the patient has been exposed to mold; contacting the tricothecene mycotoxin with an antibody directed against the tricothecene mycotoxin to quantify the tricothecene mycotoxin, wherein the antibody detects macrocyclic tricothecenes; and quantifying the tricothecene mycotoxin to determine whether the patient has a fungal infection that produces macrocuclic tricothecenes wherein the sensitivity of the quantitation is 0.2 ng/ml.
2. 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.
3. A method of determining if a patient is at risk for or has developed a disease state related to a fungal infection that produces macrocyclic tricothecenes, the method comprising the steps of: extracting and recovering a macrocyclic tricothecene mycotoxin from a tissue or body fluid of the patient wherein the body fluid is selected from the group consisting of urine sample, a nasal secretion sample, a nasal wash sample, and a sputum sample wherein the patient has been exposed to mold; contacting the tricothecene mycotoxin with an antibody directed against the tricothecen mycotoxin, wherein the antibody detects macrocyclic tricothecenes; and quantifying the tricothecene mycotoxin to determine if the patient is at risk for or has developed the disease state related to the fungal infection that produces macrocyclic tricothecenes wherein the sensitivity of quantitation is 0.2 ng/ml.
4. The method of claim 3 further comprising the step of developing an effective treatment regimen for the patient.
5. The method of claim 3 wherein the tissue is derived from a patient tissue biopsy and is in a 10% formalin solution or is in a paraffin block.
6. The method of claim 1 , wherein the tricothecene mycotoxin is contacted with the antibody using an enzyme-linked immunosorbent assay.
7. The method of claim 3 wherein the tricothecene mycotoxin is contacted with the antibody using an enzyme-linked immunosorbent assay.
8. The method of claim 1 further comprising the step of identifying the tricothecene mycotoxin using negative and positive control samples.
9. The method of claim 3 , further comprising the step of identifying the tricothecene mycotoxin using negative and positive control samples.
10. The method of claim 1 further comprising the step of using calibration reagents to quantify the tricothecene mycotoxin.
11. The method of claim 3 further comprising the step of using calibration reagents to quantify the tricothecene mycotoxin.
12. The method of claim 1 wherein there is no detection of aflatoxins or ochratoxins.
13. The method of claim 3 wherein there is no detection of aflatoxins or ochratoxins.