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Method for counting colibacillus in water body rapidly

A technology of Escherichia coli and water body, which is applied in the field of rapid detection of Escherichia coli concentration in water body and rapid counting of Escherichia coli in water body. Short, high-sensitivity effects

Inactive Publication Date: 2007-07-25
EAST CHINA NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have the disadvantages of long detection cycle, complicated procedures, and many reagents required, and it is difficult to meet the requirements of rapid detection of E. coli concentration.

Method used

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  • Method for counting colibacillus in water body rapidly

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Effect test

Embodiment 1

[0027] In the first step, the modification solution is IrCl 3 , PdCl 2 、Na 2 SO 4 Mixed liquid with water, capacity is 50mL, in this modified liquid, IrCl 3 , PdCl 2 and Na 2 SO 4 The contents of the samples were 0.005mmol, 0.0025mmol and 0.005mmol, respectively, and the balance was water. The scanning speed was 0.05V / s, and the scanning cycle was 45 cycles. In the second step, 1.0 mg of isopropyl β-D-thiogalactopyranoside was added to each culture flask, shake cultured at 45°C for 3 hours, and 0.1 mg of polymyxin B was added to each culture flask. Peptide and 0.25 mg lysozyme were shaken at 40°C for 40 minutes, and then 6.0 mg of 4-aminophenol β-D-galactopyranose was added to each flask, and incubated in a water bath at 50°C for 40 minutes. In the third step, the pH value of the phosphate buffer solution is 6.0, the content of methanol is 10%, and the flow rate is 0.6mL / min. The results obtained by detecting E. coli solutions of different known concentrations are shown...

Embodiment 2

[0031] In the first step, the modification solution is IrCl 3 , PdCl 2 、Na 2 SO 4 Mixed liquid with water, capacity is 50mL, in this modified liquid, IrCl 3 , PdCl 2 and Na 2 SO 4 The contents are 0.01mmol, 0.005mmol and 0.01mmol respectively, the balance is water, the scanning speed is 0.1V / s, and the scanning cycle is 30 circles; in the second step, 2.0mg of isopropyl beta is added to each culture bottle -D-thiogalactoside, cultured with shaking at 37°C for 2 hours, added 0.1 mg of polymyxin B nonapeptide and 0.25 mg of lysozyme to each flask, shaken at 30°C for 30 minutes, and added to each flask Add 8.0 mg of 4-aminophenol β-D-galactopyranose to the culture bottle, and incubate in a water bath at 44.5°C for 25 minutes; in the third step, the pH value of the phosphate buffer solution is 7.0, the content of methanol is 20%, and the flow rate is 0.8mL / min, the results of detecting E. coli solutions with different known concentrations are shown in Table 2. In the fitted...

Embodiment 3

[0035] In the first step, the modification solution is IrCl 3 , PdCl 2 、Na 2 SO 4 Mixed liquid with water, capacity is 50mL, in this modified liquid, IrCl 3 , PdCl 2 and Na 2 SO 4 The contents are 0.015mmol, 0.01mmol and 0.015mmol respectively, the balance is water, and the scanning speed is 0.15V / s, and the scanning cycle is 15 circles; in the second step, 3.0mg of isopropyl beta is added to each culture bottle -D-Thiogalactoside, shaking culture at 30°C for 1 hour, adding 0.1mg of polymyxin B nonapeptide and 0.25mg of lysozyme to each flask, shaking at 20°C for 20 minutes, and then adding to each flask Add 10.0 mg of 4-aminophenol β-D-galactopyranose to the culture bottle, and incubate in a water bath at 40°C for 15 minutes; in the third step, the pH value of the phosphate buffer solution is 8.0, the content of methanol is 30%, and the flow rate is 1.0mL / min, the results of detecting E. coli solutions with different known concentrations are shown in Table 3. In the fi...

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Abstract

This invention discloses a method for rapidly detecting E. coli concentration in water body. The method comprises: utilizing IrO2 / Pd decorative electrode as the electrochemical detector for flow injection-Ampere analysis, adding inducer isopropyl beta-D-thiogalactoside (IPTG), penetrant polymixin B nonapeptide and lysozyme into E. coli solution, releasing beta-D-galactosidase from E. coli cells induced by IPTG and catalytically hydrolyzing substrate 4-aminophenol-beta-D-galactopyranose in E. coli solution to obtain 4-aminophenol. Since the E. coli concentration in E. coli solution is in linear relationship with the amount of 4-aminophenol, rapidly detecting the amount of 4-aminophenol can realize rapid detection of E. coli concentration. The method has such advantages as short detection time, easy operation and high detection accuracy.

Description

technical field [0001] The present invention relates to a kind of method for quickly counting coliform bacteria in water body, promptly a kind of method for fast detection coliform bacteria concentration in water body, exactly, relate to a kind of method that uses flow injection-IrO 2 The invention discloses a method for rapidly detecting the concentration of Escherichia coli in water by amperometric analysis of a Pd modified electrode, which belongs to the technical field of microbiological inspection of water. Background technique [0002] The microbiological examination of water body, especially the detection of the concentration of intestinal bacteria is of great significance in ensuring the safety of drinking water and controlling infectious diseases. The concentration of E. coli in water sources is an important indicator that directly reflects the pollution of water sources by human and animal excrement. Therefore, the concentration of E. coli in drinking water, surfac...

Claims

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Application Information

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IPC IPC(8): C12Q1/10
Inventor 张文耿萍唐辉张新爱郑娇红朱伟王丹安雅睿金利通
Owner EAST CHINA NORMAL UNIV
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