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Substrate combination, detection reagent and detection kit for detecting escherichia coli and/or shigella

A technology for Escherichia coli and Shigella, which is applied to measurement devices, material analysis, instruments and other directions by observing the impact on chemical indicators, can solve the problems of inability to identify Shigella, time-consuming, and complicated steps.

Active Publication Date: 2021-01-01
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Patent CN110144379A adopts a special treatment method. It is necessary to carry out differential culture of the components of the sample to be tested for 2h-24h in advance to achieve strain-specific protein expression. The whole operation process involves recultivation, mass spectrometry pretreatment, and mass spectrometry secondary identification, and the steps are cumbersome and time-consuming
[0011] Patent CN110144379A also has a serious deficiency: in this technical solution, only Escherichia coli and Shigella can be identified and distinguished. If it is Shigella, it can only be identified at the genus level, but Shigella cannot be identified. Species of the following genus

Method used

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  • Substrate combination, detection reagent and detection kit for detecting escherichia coli and/or shigella
  • Substrate combination, detection reagent and detection kit for detecting escherichia coli and/or shigella
  • Substrate combination, detection reagent and detection kit for detecting escherichia coli and/or shigella

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] A clinical strain, the result of mass spectrometry identification is Escherichia coli, the operation steps are as follows:

[0071] 1. Board preparation, as follows:

[0072] Well 1: 0.8% mannitol, 1.4% phenol red, phosphate buffer pH7.2

[0073] Well 2: 0.8% ONPG (2-nitrophenyl-β-D-galactopyranoside), phosphate buffer pH7.0

[0074] Well 3: 0.8% sorbitol, 1.4% phenol red, phosphate buffer pH7.2

[0075] Well 4: 0.5% glycerol, 1.4% phenol red, phosphate buffer pH7.2

[0076] 12ul of each coating solution was coated on a four-well plate card and used after natural drying

[0077] 2. Preparation of bacterial suspension: use a turbidimeter to prepare 2.5 liters of Maibella bacteria from the strain to be tested, add 100ul per well to the above-mentioned plate, and place the plate in a 37°C incubator for 1.5h after adding the sample.

[0078] 3. Interpretation of results: Take out the board, record the color of the reaction well and the corresponding yin and yang results...

Embodiment 2

[0086] A clinical strain, the result of mass spectrometry identification is Escherichia coli, the operation steps are as follows:

[0087] 1. Board preparation, as follows:

[0088] Well 1: 1.2% mannitol, 1.7% phenol red, Tris buffer pH7.4

[0089] Well 2: 1.2% ONPG (2-nitrophenyl-β-D-galactopyranoside), Tris buffer pH8.0

[0090] Well 3: 1.2% sorbitol, 1.7% phenol red, Tris buffer pH7.4

[0091] Well 4: 1% glycerol, 1.7% phenol red, Tris buffer pH7.4

[0092] 8ul of each coating solution was coated on a four-well plate, and used after natural drying

[0093] 2. Preparation of bacterial suspension: use a turbidimeter to prepare 3.0 strains of Maibella bacteria from the strain to be tested, add 100ul per well to the above-mentioned plate, and place the plate in a 37°C incubator for 1 hour after adding the sample.

[0094] 3. Interpretation of results: Take out the board, record the color of the reaction well and the corresponding yin and yang results

[0095] Well 1: yello...

Embodiment 3

[0102] A clinical strain, the result of mass spectrometry identification is Escherichia coli, the operation steps are as follows:

[0103] 1. Board preparation, as follows:

[0104] Well 1: 1% mannitol, 1.6% phenol red, phosphate buffer pH7.3

[0105] Well 2: 1% ONPG (2-nitrophenyl-β-D-galactopyranoside), phosphate buffer pH7.5

[0106] Well 3: 1% sorbitol, 1.6% phenol red, phosphate buffer pH7.3

[0107] Well 4: 0.8% glycerol, 1.6% phenol red, phosphate buffer pH7.3

[0108] 10ul of each coating solution was coated on a four-well plate card, and used after natural drying

[0109] 2. Preparation of bacterial suspension: use a turbidimeter to prepare 2.8 molasses strains from the strain to be tested, add 100ul per well to the above-mentioned plate, and place the plate in a 37°C incubator for 2 hours after adding the sample.

[0110] 3. Interpretation of results: Take out the board, record the color of the reaction well and the corresponding yin and yang results

[0111] Ho...

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PUM

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Abstract

The invention relates to the field of biological detection, and particularly relates to a substrate combination, a detection reagent and a detection kit for detecting escherichia coli and / or shigella.The substrate combination provided by the invention is a detection kit for quickly distinguishing escherichia coli and the shigella based on biochemical and chromogenic principles and a preparation method thereof is provided. By using the kit disclosed by the invention, the escherichia coli, shigella bogdii, shigella sonnei, shigella flexneri and shigella dysenteriae can be quickly distinguishedwithin 1-2 hours by only one step of operation.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a substrate combination, a detection reagent and a detection kit for detecting Escherichia coli and / or Shigella. Background technique [0002] Microbiological identification is an important part of food, drug, customs and clinical inspection. The main methods include: [0003] (1) Biochemical identification based on biochemical reaction spectrum: because biochemical identification involves more biochemical reactions, a biochemical identification system has been developed. Based on bacterial culture and different biochemical profiles of different bacteria to achieve species identification, the identification results are accurate and can be It is accurate to species or even subspecies, but the identification of bacterial species and speed is not as good as sequencing technology and mass spectrometry technology. [0004] (2) Sequencing technology based on 16sRNA sequencing: Seq...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N27/62G01N33/569
CPCG01N21/78G01N33/6848G01N33/56916G01N2021/7759Y02A50/30
Inventor 崔晓莉郑业焕方娟吴小瑞李晓帅付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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