Method for preparing a measurement sample for detecting live cells of a microorganism in a test sample
AssigneeMorinaga Milk Industry Co., Ltd.
InventorsShinichi Yoshida, Takashi Soejima
Life of this patent
Estimating the term…
Abstract
According to the following steps, live cells, injured cells, VNC cells and dead cells of a microorganism in a test sample are detected by flow cytometry: a) the step of treating the test sample with an enzyme having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample, b) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison. c) the step of treating the test sample treated in the steps a) and b) with a nuclear stain agent, and d) the step of detecting the microorganism in the test sample treated with the nuclear stain agent by flow cytometry.
Claims5
A method for preparing a measurement sample for detecting live cells of a microorganism in a test sample by flow cytometry, which comprises the following steps:
- a) treating the test sample with an enzyme comprising lipolytic enzymes and proteases and having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample, and
- b) treating the test sample with a topoisomerase poison selected from ethidium monoazide, amsacrine, camptothecin, doxorubicin, ellipticine, etoposide, mitoxantrone, saintopin, topotecan and CP-115,953, and/or a DNA gyrase poison selected from ciprofloxacin, ofloxacin, enoxacin, pefloxacin, fleroxacin, norfloxacin, nalidixic acid, oxolinic acid and piromidic acid, and
- c) treating the test sample treated in steps a) and b) with propidium iodide and SYTO9.
Dependencies are inferred from the claim text.