Method for detection of microorganism and kit for detection of microorganism

摘要

Live cells of a microorganism in a test sample are detected by the following steps:
a) the step of treating the test sample with a topoisomerase poison and/or a DNA gyrase poison,b) the step of extracting DNA from the test sample, and amplifying a target region of the extracted DNA by PCR, andc) the step of analyzing an amplification product.

主权项

1. A method for detecting live cells by distinguishing the live cells from dead cells and also from injured cells of a microorganism in a test sample, wherein the microorganism is Listeria bacteria, which comprises the following steps:
a) treating the test sample with ethidium monoazide and irradiating the test sample with visible light; a′) treating the test sample with a topoisomerase poison other than ethidium monoazide and/or a DNA gyrase poison other than ethidium monoazide, wherein the topoisomerase poison other than ethidium monoazide and/or a DNA gyrase poison other than ethidium monoazide is selected from the group consisting of amsacrine of a final concentration of 1 to 100 μg/ml, ellipticine of a final concentration of 0.05 to 5 μg/ml, camptothecin of a final concentration of 1 to 100 μg/ml, ciprofloxacin of a final concentration of 0.4 to 40 μg/ml, etoposide of a final concentration of 1 to 100 μg/ml, and mitoxantrone of a final concentration of 0.1 to 10 μg/ml; b) extracting DNA from the test sample and amplifying a target region of the extracted DNA by PCR; c) analyzing a resulting PCR amplification product; and d) distinguishing the live cells from dead cells and also from injured cells.