Methods for detecting microbial growth, methods for obtaining microorganisms, kits for detecting microbial growth, kits for obtaining microorganisms, and use of dyes as microbial growth reporters.

Abstract

To provide a method for detecting the growth of microorganisms that can accurately detect a droplet in which microorganisms have grown, to provide a method for obtaining microorganisms, to provide a kit for detecting the growth of microorganisms, to provide a kit for obtaining microorganisms, and to provide use of dye as microbial growth reporter.SOLUTION: A method for detecting the growth of microorganisms comprises: producing a droplet containing a dye whose fluorescence intensity changes by interacting with a microorganism and a membrane component possessed by the microorganism, in W / O emulsion; culturing the microorganism within the droplet; and detecting the growth of the microorganism based on the fluorescence intensity of the dye.SELECTED DRAWING: Figure 1

Claims

1. To prepare droplets containing microorganisms and styryl dye in a W / O emulsion, The microorganisms are cultured in the droplet, Based on the fluorescence intensity of the styryl dye, the growth of the microorganism is detected with a lower limit of 10 cells as the average number of cells per droplet. Includes, The aforementioned styryl dye is N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide or 4-[6-[4-(diethylamino)phenyl]-1,3,5-hexatrien-1-yl]-1-[3-(triethylammonio)propyl]-pyridinium dibromide Method for detecting microbial growth.

2. The aforementioned microorganisms There are multiple species. The method for detecting microbial growth according to claim 1.

3. To prepare droplets containing microorganisms and styryl dye in a W / O emulsion, The microorganisms are cultured in the droplet, Based on the fluorescence intensity of the styryl dye, the growth of the microorganism is detected with a lower limit of 10 cells as the average number of cells per droplet. The droplets in which the growth of the aforementioned microorganisms was detected are collected, Includes, The aforementioned styryl dye is N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide or 4-[6-[4-(diethylamino)phenyl]-1,3,5-hexatrien-1-yl]-1-[3-(triethylammonio)propyl]-pyridinium dibromide How to obtain microorganisms.

4. A compound comprising N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide or 4-[6-[4-(diethylamino)phenyl]-1,3,5-hexatrien-1-yl]-1-[3-(triethylammonio)propyl]-pyridinium dibromide A kit for detecting microbial growth in droplets in a W / O emulsion, for use in the microbial growth detection method described in claim 1 or 2.

5. A compound comprising N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide or 4-[6-[4-(diethylamino)phenyl]-1,3,5-hexatrien-1-yl]-1-[3-(triethylammonio)propyl]-pyridinium dibromide A kit for acquiring microorganisms in droplets in a W / O emulsion for use in the method for acquiring microorganisms described in claim 3.

6. The use of styryl dye as a microbial growth reporter in droplets in a W / O emulsion, The lower limit of the average number of cells contained in one droplet detected by the microbial growth reporter is 10 cells. The aforementioned styryl dye is N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide or 4-[6-[4-(diethylamino)phenyl]-1,3,5-hexatrien-1-yl]-1-[3-(triethylammonio)propyl]-pyridinium dibromide use.

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