Rapid detection of mold by accelerated growth and detection

摘要

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.

主权项

1. A method of detecting the presence or absence of a mold having hyphae, comprising:
(a) applying a sample suspected of containing the mold to a volume of liquid growth medium containing a porous substrate attached to a shaft, wherein said porous substrate attached to a shaft is not used to collect the sample prior to step a); (b) allowing growth of said mold to occur in said liquid growth medium of step a) and for said mold to attach onto said porous substrate attached to a shaft; (c) transferring said mold attached to said porous substrate attached to a shaft from step b) to a phosphate buffer solution by:
i) removing said porous substrate attached to a shaft with attached mold from said liquid growth medium; andii) placing said porous substrate attached to a shaft with attached mold into a smaller volume of phosphate buffer solution, wherein said volume of phosphate buffer solution is less than the volume of liquid growth medium from which it was removed; d) mechanically fragmenting said mold in said volume of phosphate buffer solution from step c) by subjecting said porous substrate attached to a shaft with attached mold to a shear force to disrupt connections between segments of the hyphae and produce fragmented mold hyphae segments; e) associating said fragmented mold hyphae segments sample of step d) with a labeling agent; and f) directly detecting the presence or absence of said mold hyphae segments with said labeling agent of step e) by flow cytometry;
wherein the liquid growth medium comprises a nitrogenous substance and a carbon source.