Methods and compositions for identifying yeast
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.
1. A method of identifying a specific yeast species in patient tissue or body fluid, the method comprising 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, wherein the probe sequence is selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 10, a full complement of the sequence of SEQ ID NO: 4, and a full complement of the sequence of SEQ ID NO: 10; and
specifically identifying the yeast species, wherein the yeast species is Candida glabrata or Candida tropicalis.
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 body fluids are selected 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, and plasma.
4. The method of claim 1 wherein the DNA is amplified using PCR.
5. The method of claim 4 wherein the PCR is real-time PCR.
6. The method of claim 1 wherein the probe is fluorescently labeled.
7. The method of claim 5 wherein the probe is fluorescently labeled.
8. The method of claim 2 wherein the probe, a forward primer, and a reverse primer are used during the amplification step and the probe comprises the sequence of SEQ ID NO: 4, the forward primer comprises the sequence of SEQ ID NO: 5, and the reverse primer comprises the sequence of SEQ ID NO: 6.
9. The method of claim 2 wherein the probe, a forward primer, and a reverse primer are used during the amplification step and the probe comprises the sequence of SEQ ID NO: 10, the forward primer comprises the sequence of SEQ ID NO: 11, and the reverse primer comprises the sequence of SEQ ID NO: 12.
10. The method of claim 2 wherein the amplified sequence is internal transcribed spacer regions of nuclear ribosomal DNA.
11. The method of claim 1 wherein the probe is bound to a bead dyed with a fluorochrome.
12. The method of claim 1 further comprising hybridizing a probe to the DNA to specifically identify other yeast species, wherein said probe has a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 7, a full complement of the sequence of SEQ ID NO: 1, and a full complement of the sequence of SEQ ID NO: 7.
13. The method of claim 12 wherein the amplifying step is performed with primers that hybridize to the DNA.
14. The method of claim 13 wherein the probe, a forward primer, and a reverse primer are used during the amplification step and the probe comprises the sequence of SEQ ID NO: 1, the forward primer comprises the sequence of SEQ ID NO: 2, and the reverse primer comprises the sequence of SEQ ID NO: 3.
15. The method of claim 13 wherein the probe, a forward primer, and a reverse primer are used during the amplification step and the probe comprises the sequence of SEQ ID NO: 7, the forward primer comprises the sequence of SEQ ID NO: 8, and the reverse primer comprises the sequence of SEQ ID NO: 9.
16. The method of claim 12 wherein the other yeast species are selected from the group consisting of Candida albicans and Candida kruseii.
17. The method of claim 12 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 12 wherein the probe is fluorescently labeled.