Method for detecting fluorine content in escherichia coli

A technology of Escherichia coli and fluorine content, applied in the field of microbial detection, can solve the problem of inability to separately determine the effective concentration of fluorine and lethal components in drugs, avoid excessive fluorine residues, make the ratio reasonable and efficient, and increase permeability. sexual effect

Active Publication Date: 2020-12-04
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Conventional drug susceptibility tests can only determine the effective concentration range of antibacterial drugs, and cannot separately determine the effective concentrations of fluorine and lethal components in the drug

Method used

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  • Method for detecting fluorine content in escherichia coli
  • Method for detecting fluorine content in escherichia coli
  • Method for detecting fluorine content in escherichia coli

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1: the mensuration of escherichia coli growth OD600 value

[0059] 1) Preparation of LB liquid medium: 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride were added to 1000mL deionized water and mixed thoroughly; 800mM NaF mother liquor was prepared; 200mL liquid LB was weighed respectively, and NaF mother liquor was added to it to prepare Prepare fluorine-containing LB liquid medium with fluoride ion concentrations of 0mM, 15mM, 25mM, and 50mM, adjust the pH to 7.0, sterilize at 120°C for 20min, and cool to 40°C;

[0060] 2) Add 2 mL of Escherichia coli wild-type strain bacterial liquid to the fluorine-containing liquid LB in step 1), the ratio of the bacterial liquid to the fluorine-containing liquid LB is 1:100, and shake at a constant temperature of 37 ° C and 220 rpm for 10 h;

[0061] 3) Using a UV spectrophotometer (MAPADA P4) at a wavelength of 600nm to detect the OD600 values ​​of Escherichia coli growth in 0mM, 15mM, 25mM, and 50mM flu...

Embodiment 2

[0062] Embodiment 2: Determination and calculation of Escherichia coli fluoride accumulation

[0063] 1) Collect the cultured bacterial solution, divide it into 50mL centrifuge tubes, and centrifuge at 25°C and 4000r / min for 15min;

[0064] 2) Pour off the supernatant after centrifugation, add 30mL deionized water to resuspend the bacterial mass at the bottom, and then centrifuge at 25°C and 4000r / min for 15min;

[0065] 3) Repeat step 2) twice;

[0066] 4) Pour off the supernatant after centrifugation in step 3), add 30mL deionized water, resuspend, heat in a water bath at 100°C for 30min, and after cooling to room temperature, centrifuge at 25°C and 4000r / min for 15min;

[0067] 5) Take the centrifuged supernatant in step 4), add 15mL of fluoride ion strength buffer at a ratio of 1:1, mix thoroughly, and measure with a fluoride ion selective electrode (Thermo Orin 9609BNWP) to obtain the fluoride ion concentration C;

[0068] 6) Substitute the obtained data into the formu...

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Abstract

The invention belongs to the field of microbiological detection, and relates to a method for detecting fluorine content in escherichia coli, which comprises the following steps of 1) preparing LB liquid culture medium containing fluorine with different concentrations, 2) adding a wild escherichia coli strain into the LB liquid culture medium, and carrying out shaking culture, 3) carrying out ultraviolet detection on the OD600 value of the growth of the wild escherichia coli, 4) collecting the cultured bacterial liquid, centrifuging, pouring out the supernatant, re-suspending, re-centrifuging,pouring out the supernatant, re-suspending, heating in a water bath, cooling and centrifuging, 5) taking the supernatant, and measuring by using a fluorine ion selective electrode to obtain fluorine ion concentration C, and 6) calculating to obtain the fluorine content accumulated in the wild escherichia coli.The invention provides a method for detecting the fluorine content in escherichia coli. The method provides concentration reference and proportion basis for research and development of fluorine-containing antibacterial drugs, and solves the problem that the effective concentrations of fluorine and lethal components in the drugs cannot be determined respectively during research and development of the fluorine-containing antibacterial drugs.

Description

technical field [0001] The invention belongs to the field of microorganism detection, and relates to a method for detecting fluorine content in Escherichia coli. Background technique [0002] Escherichia coli is an opportunistic pathogenic bacterium that can live in the intestines of humans or animals under suitable conditions and easily cause gastrointestinal infections and other diseases. The occurrence and prevalence of Escherichia coli infection are distributed worldwide and have the characteristics of regional distribution, especially sick or infected animals often become the source of food contamination from animal sources, such as beef or dairy products from infected cattle, from Eggs or chicken products from infected chickens. In addition, sick or infected animals can also pollute local pastures and water sources through excreted feces, resulting in cross-contamination and even regional outbreaks of E. coli. Mild symptoms of E. coli infection usually only cause dia...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/333G01N21/33
CPCG01N27/26G01N27/333G01N21/33
Inventor 王玉花武子辰邢安琪徐晓寒田志强房婉萍朱旭君马媛春
Owner NANJING AGRICULTURAL UNIVERSITY
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