Method for determining escherichia coli content in water

A technology of Escherichia coli and content, applied in the field of microbial detection, can solve the problems that ELISA detection results are easily affected by various factors, difficult to meet rapid detection, and prone to false positives, etc., and achieve the goal of avoiding intrinsic Raman signals and fluorescent signals Effects of interference, shortened detection time, and good optical stability

Pending Publication Date: 2020-06-26
WUHAN ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The traditional detection and enumeration method of Escherichia coli is the MPN method (most probable number, maximum probable number), the basis of this method is that Escherichia coli carries out lactose fermentation, gas production and indole formation on the growth medium, but this method requires 6 -7 days, can no longer adapt to the principles of modern quality management
[0004] Current new amplification techniques, including immunoassay (enzyme-linked immunosorbent assay (ELISA)) and nucleic acid identification (polymerase chain reaction (PCR)), etc., all require tedious procedures, and the detection results of ELISA are susceptible to many Affected by various factors, false positives are prone to occur, and it is not suitable for on-site detection; high-throughput sequencing requires the isolation of bacterial DNA in advance, in addition to professional operators, and expensive nucleic acid amplification equipment, which is difficult to meet the urgent need for rapid on-site detection need

Method used

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  • Method for determining escherichia coli content in water
  • Method for determining escherichia coli content in water
  • Method for determining escherichia coli content in water

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment

[0038] Establishment of standard working curve

[0039] (1) Instruments and reagents

[0040] Portable Raman spectrometer with excitation wavelength of 785nm;

[0041] Silver-Escherichia coli system: self-synthesized by hydroxylamine hydrochloride reduction method, its transmission electron microscope is as follows image 3 ;

[0042] Capture probe: E. coli monoclonal antibody modified on the surface of carboxyl magnetic beads, self-prepared, and its transmission electron microscope is such as Figure 4 ;

[0043] Gold nanosol: self-synthesized by sodium citrate reduction method of chloroauric acid;

[0044] Detection probe: MBN-modified gold nanoparticles, self-prepared;

[0045] Different concentrations of Escherichia coli bacteria, self-cultivation.

[0046] (2) Experimental steps

[0047] In situ reduction growth of silver nanomaterials on the surface of Escherichia coli with different concentrations, after mixing with capture probes, they were separated under the a...

Embodiment 1

[0049] Take 500ml of ultrapure water, add different concentrations of Escherichia coli to it, and operate according to the steps and detection methods described above. The results are shown in Table 1.

Embodiment 2

[0051] Take 5000ml of tap water, add different concentrations of Escherichia coli into it respectively, and operate according to the steps and detection methods described above. The results are shown in Table 1.

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Abstract

The invention relates to the technical field of microbiological detection, and provides a method for determining the content of escherichia coli in water. The method comprises the following steps: preparation of a silver-escherichia coli system, preparation of a capture probe, addition of nitric acid, preparation of a detection probe, establishment of an escherichia coli detection standard workingcurve, and determination of the content of escherichia coli in water. The invention provides an SERS detection method of escherichia coli. The SERS detection method can be used for specifically, sensitively and quickly detecting the content of escherichia coli in water.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular relates to a method for determining the content of Escherichia coli in water quality. Background technique [0002] Escherichia coli (E.coli for short) is habitually called Escherichia coli. It is a normal resident bacteria in the intestinal tract of humans and various animals. Therefore, once E. coli is detected, it means that the sample is directly or indirectly contaminated with feces. Therefore, Escherichia coli is used as a hygienic bacteriological indicator of fecal-derived contamination of drinking water, food, and the like. The detection limit of Escherichia coli is clearly stipulated in the relevant national standards for food and drinking water in my country. Among them, the standard for Escherichia coli in drinking water in my country's "Sanitary Standards for Drinking Water" GB5749-85 stipulates that 1L of water does not exceed 3 If only chlorinated ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65C12N1/20C12N1/02C12R1/19
CPCG01N21/65G01N21/658C12N1/20C12N1/02
Inventor 白向茹王利华肖康飞韩艳云
Owner WUHAN ACADEMY OF AGRI SCI
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