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 identifying a specific fungal species in patient tissue or body fluid, the method comprising the steps of:
extracting and recovering DNA of the fungal species from the patient tissue or body fluid;
amplifying the DNA;
hybridizing an isolated probe to the DNA to specifically identify the fungal species, wherein the probe consists of a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 40, and SEQ ID NO: 67, and wherein the probe is fluorescently labeled; and
specifically identifying the fungal species.
2. The method of claim 1 wherein the amplifying step is performed with primers that hybridize to the DNA.
3. The method of claim 1 wherein the DNA is amplified using PCR.
4. The method of claim 3 wherein the PCR is real-time PCR.
5. The method of claim 2 wherein the probe is the sequence consisting of SEQ ID NO: 1.
6. The method of claim 2 wherein the probe is the sequence consisting of SEQ ID NO: 40.
7. The method of claim 2 wherein the probe is the sequence consisting of SEQ ID NO: 67.
8. The method of claim 2 wherein the amplified sequence is internal transcribed spacer regions of nuclear ribosomal DNA.
9. The method of claim 1 wherein the probe is bound to a bead dyed with a fluorochrome.
10. A method of determining if a patient is at risk for or has developed a disease state related to a fungal infection, the method comprising the steps of:
extracting and recovering DNA of a specific fungal species from a tissue or body fluid of the patient;
amplifying the DNA;
hybridizing a probe to the DNA to specifically identify the fungal species, wherein the probe consists of a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 40, and SEQ ID NO: 67, and wherein the probe is fluorescently labeled; and
specifically identifying the fungal species.
11. The method of claim 10 further comprising the step of developing an effective treatment regimen for the patient.
12. The method of claim 10 wherein the probe is bound to a bead dyed with a fluorochrome.
13. The method of claim 10 wherein the probe is the sequence consisting of SEQ ID NO: 1.
14. The method of claim 10 wherein the probe is the sequence consisting of SEQ ID NO: 40.
15. The method of claim 10 wherein the probe is the sequence consisting of SEQ ID NO: 67.
16. The method of claim 10 wherein the amplifying step is performed with primers that hybridize to the DNA to produce an amplified sequence.
17. The method of claim 10 wherein the DNA is amplified using PCR.
18. The method of claim 17 wherein the PCR is real-time PCR.
19. The method of claim 16 wherein the amplified sequence is internal transcribed spacer regions of nuclear ribosomal DNA.