Method and kit for detection of microorganism

摘要

Live cells of microorganism in a test sample are detected by distinguishing the live cells from dead cells or injured cells by the following steps of:
a) adding an agent capable of covalently binding to DNA or RNA by irradiation with light having a wavelength of 350 nm to 700 nm to the test sample;b) irradiating the test sample to which the cross-linking agent is added with light having a wavelength of 350 nm to 700 nm;c) amplifying a target region of DNA or RNA of the microorganism contained in the test sample by a nucleic acid amplification method in the presence of an agent for suppressing an action of a nucleic acid amplification inhibitory substance, without extracting nucleic acids from the cells; andd) analyzing the amplified product.

主权项

1. A method for detecting live cells of a microorganism in a test sample by distinguishing the live cells from dead cells or injured cells, which comprises the steps of:
a) adding an agent capable of covalently binding to DNA or RNA by irradiation with light having a wavelength of 350 nm to 700 nm to the test sample; b) irradiating the test sample to which the agent is added with light having a wavelength of 350 nm to 700 nm; c) amplifying a target region of DNA or RNA of the microorganism contained in the test sample by a nucleic acid amplification method in the presence of
(i) lysozyme and/or a nonionic surfactant,(ii) a magnesium salt,(iii) either one of an organic acid salt and a phosphoric acid salt, and(iv) one or more selected from the group consisting of albumin, dextran, T4 gene 32 protein, acetamide, betaine, dimethyl sulfoxide, formamide, glycerol, polyethylene glycol soybean trypsin inhibitor, α2-macroglobulin, tetramethylammonium chloride, lysozyme, phosphorylase and lactate dehydrogenase without extracting nucleic acids from the cells, and d) analyzing the amplified product, thereby detecting the live cells.