Rapid detection of mold by accelerated growth and detection

Abstract

The invention includes a method of detecting the presence or absence of mold in a sample. The method can include the step of applying a sample suspected of containing the mold to a growth medium, optionally fragmenting the sample, associating the sample with a labeling agent, and detecting the presence or absence of the mold by detection of the labeling agent. The invention can also include a medium for growing mold that includes a stimulation agent.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a scatter plot of mold hyphae stained with a fluorochrome in a growth medium in accordance with an embodiment of the invention.

FIG. 2 shows a scatter plot of the growth medium of FIG. 1 without mold hyphae in accordance with an embodiment of the invention.

Claims

1. A method of detecting the presence or absence of a mold, comprising: applying a sample suspected of containing the mold to a growth medium;fragmenting the sample;associating the sample with a labeling agent; anddetecting the presence or absence of the mold in the sample by detecting the labeling agent.

2. The method of claim 1, wherein the sample includes a personal care product.

3. The method of claim 1, wherein the sample includes an over-the-counter product.

4. The method of claim 1, wherein the labeling agent includes a fluorescent agent.

5. The method of claim 4, wherein the fluorescent agent is selected from the group consisting of nucleic acid dyes, nucleic acid stains, fluorochrome labeled anti-mold antibodies, mold specific stains, fluorochrome labeled nucleic acid probes, and combinations thereof.

6. The method of claim 4, wherein the fluorescent agent includes fluorophores immobilized on a surface.

7. The method of claim 6, wherein the surface is selected from the group consisting of latex spheres, magnetized spheres, and nanoparticles.

8. The method of claim 4, further including the step of adding a quenching agent after associating the sample with the fluorescent agent.

9. The method of claim 8, wherein the quenching agent comprises Acid Black.

10. The method of claim 1, wherein the growth medium comprises a nitrogenous substance, a sample neutralization agent, a carbon source, and a stimulation agent.

11. The method of claim 10, wherein the stimulation agent includes n-acetyl glucosamine.

12. The method of claim 1, wherein a solid article is included with the growth medium, further comprising the step of removing the solid article from the growth medium.

13. The method of claim 12, further including the step of placing the solid article in a buffer solution.

14. The method of claim 1, wherein the sample is fragmented chemically by a chemical fragmenting agent.

15. The method of claim 14, wherein the chemical fragmenting agent comprises an agent selected from the group consisting of organic acids, inorganic acids, inorganic bases, oxidizing agents, and enzymes that attack cell walls.

16. The method of claim 14, wherein the chemical fragmenting agent comprises acetic acid.

17. The method of claim 1, wherein the sample is fragmented mechanically.

18. The method of claim 17, wherein the mechanically fragmenting step includes one or more steps from the group consisting of subjecting the sample to sonic energy, high shear force, and mechanical agitation.

19. The method of claim 1, wherein the detecting step includes placing the sample in a flow cytometer.

20. The method of claim 19, wherein the labeling agent includes a nucleic acid dyes/stain selected from the group consisting of Syto 62, Syto 61, Syto 59, and Hexidium Iodide.

21. The method of claim 20, wherein the labeling agent includes Hexidium Iodide.

22. The method of claim 1, wherein the detecting step includes placing the sample in a fluorescent sample chamber.

23. The method of claim 19, wherein the flow cytometer produces a scatter plot.

24. The method of claim 23, wherein the scatter plot is compared to a scatter plot of a known mold-free sample to determine if the sample contains mold.

25. The method of claim 1, wherein the labeling agent includes a fluorescent agent and a total fluorescence is detected.

26. The method of claim 1, wherein the presence or absence of the mold can be detected within 24 hours of the applying step.

27. A medium for growing mold, the medium comprising, a nitrogenous substance;a sample neutralization agent;a carbohydrate source;a carbon source; anda stimulation agent.

28. The medium of claim 27, wherein the nitrogenous substance includes Tryptic Soy Broth.

29. The medium of claim 27, wherein the neutralization agent includes soy lecithin and/or Tween-20.

30. The medium of claim 27, wherein the carbohydrate source includes malt extract.

31. The medium of claim 27, wherein the carbon source includes glucose.

32. The medium of claim 27, wherein the stimulation agent includes n-acetyl glucosamine.

33. The medium of claim 27, wherein the growth medium includes one or more solid articles.

34. The medium of claim 33, wherein the solid articles are selected from the group consisting of swabs, sponges, mesh material, small bead-like objects, metallic beads, ceramic beads, chips of ceramic materials, and chips of metallic materials.

35. The medium of claim 27, further comprising a substrate useful for concentrating and isolating the sample in contact with the growth medium.

36. The medium of claim 35, wherein the substrate is porous.

37. The medium of claim 35, wherein the substrate comprises at least one material selected from the group consisting of a polyester, a polyurethane, a cotton, a foam, a rayon, a polyether, a metal or a plastic.

38. The medium of claim 35, wherein the substrate is selected from the group consisting of a swab, a sponge, a mesh screen, and a porous material.

39. The medium of claim 35, wherein the substrate is attached to a shaft useful for manipulating the substrate.

40. The medium of claim 39, wherein the shaft includes at least one material selected from the group consisting of polypropylene, wood, plastics, and metals.

41. The medium of claim 35, wherein the substrate is adhered to the bottom of a tube.

42. The medium of claim 35, wherein the substrate includes one or more surfaces derivatized to promote mold adhesion.

43. A method of detecting the presence or absence of a mold, comprising: applying a sample suspected of containing the mold to a growth medium, asubstrate useful for supporting mold growth included within the growth medium;associating the sample with a labeling agent, the labeling agent including a fluorescent agent; anddetecting the presence or absence of the mold in the sample by placing the substrate into a fluorescence chamber, the presence of mold being detectable by the labeling agent associated with the mold.

44. The method of claim 43, further including exposing the sample to a quenching agent.

45. The method of claim 44, wherein the quenching agent is added directly to the fluorescence chamber.