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Kit for detecting pathogenic bacteria and application of kit

A technology of pathogenic bacteria and kit, applied in the detection field of biological immune method

Active Publication Date: 2014-08-13
ANIMAL AND PLANT & FOOD DETECTION CENTER JIANGSU ENTRY EXIT INSPECTION AND QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are complicated to operate, have high requirements on the detection environment or instruments, and take a long time. For example, the plate method is time-consuming and has strict environmental requirements, and the instrument method is expensive and cumbersome.

Method used

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  • Kit for detecting pathogenic bacteria and application of kit
  • Kit for detecting pathogenic bacteria and application of kit
  • Kit for detecting pathogenic bacteria and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 detects the kit of Campylobacter jejuni

[0035] The kit for detecting Campylobacter jejuni includes the following reagents: gold nanoparticle solution, anti-Campylobacter jejuni antibody, apotryptophanase solution, nano-magnetic beads labeled with anti-Campylobacter jejuni antibody, bovine serum albumin solution and reactant solution . The reactant solution contains L-tryptophan and pyridoxal phosphate.

[0036] (1) Gold nanoparticles solution

[0037] The concentration of gold nanoparticles solution was 10 mM, and the average particle size of gold nanoparticles was 15 nm.

[0038] The preparation method of gold nanoparticles: add 5 mL, 1 g / L chloroauric acid solution in a conical flask, then dilute to 50 mL with sterilized water, heat to bumping under stirring, add 0.6~1 mL Sodium citrate solution with a concentration of 1% (mass percentage concentration), stirring continuously, until the color of the solution is stable to wine red.

[0039] (2) Anti-...

Embodiment 2

[0050] Embodiment 2 The present invention detects the application of Campylobacter jejuni kit

[0051] The specific steps of using the kit prepared in Example 1 to detect Campylobacter jejuni are as follows:

[0052] (1) Binding of apotryptophanase to anti-Campylobacter jejuni antibody

[0053] 1) Take 5 mL of gold nanoparticle solution, and use 0.2 mol / L K 2 CO3 Adjust the pH value of the solution to 8.0;

[0054] 2) Add 500 μL anti-Campylobacter jejuni monoclonal antibody and 500 μL apotryptophanase solution to the pH-adjusted gold nanoparticle solution, mix, shake and react at room temperature for 1 hour;

[0055] 3) Add 500 μL bovine serum albumin solution, mix, shake and react at room temperature for 30 minutes;

[0056] 4) Take out the reaction solution, centrifuge at 9.4 g for 7 minutes, take the precipitate and resuspend it in 500 μL PBST buffer to obtain the enzyme-antibody complex, store it at 4°C. After the enzyme-antibody complex is prepared, it must be used wi...

Embodiment 3

[0066] Embodiment 3 detects the kit of escherichia coli

[0067] The kit for detecting Escherichia coli is compared with the kit for detecting Campylobacter jejuni in Example 1, the only difference is that the anti-Escherichia coli monoclonal antibody is used instead of the anti-Campylobacter jejuni monoclonal antibody, and the nanomagnetic Beads replace nanomagnetic beads labeled with anti-C. jejuni monoclonal antibody.

[0068] (1) Anti-Escherichia coli monoclonal antibody

[0069] The concentration of anti-Escherichia coli monoclonal antibody is 0.1 mg / mL, purchased from Thermo Scientific Pierce, product number MA1-7029, the titer of anti-Escherichia coli monoclonal antibody is 1:20, and the affinity constant of anti-Escherichia coli monoclonal antibody is 2.8×10 8 L / M

[0070] (2) Nano-magnetic beads labeled with anti-Escherichia coli monoclonal antibody

[0071] The preparation method of nano-magnetic beads labeled with anti-Escherichia coli monoclonal antibody is as f...

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Abstract

The invention provides a kit for detecting pathogenic bacteria and application of the kit, relating to the technical field of detection of a biology immune method. The kit comprises the following reagents: gold nanoparticles, a pathogenic bacterium resisting antibody, tryptophanase, and nano magnetic beads marked by the pathogenic bacteria resisting antibody. The invention also provides application of the kit in detection of the pathogenic bacteria and a detection method. The kit for detecting the pathogenic bacteria is capable of simply, quickly and sensitively carrying out in-situ detection of the pathogenic bacteria.

Description

Technical field [0001] technical background [0002] [0003] Escherichia coli( E. Coli [0004] Invention content: [0005] [0006] [0007] [0008] The purpose of the present invention is implemented in the following technical solutions. [0009] [0010] [0011] [0012] [0013] [0014] [0015] [0016] [0017] [0018] [0019] [0020] [0021] [0022] Compared with the existing technology, the beneficial effect of the present invention is as follows: [0023] [0024] Attachment description [0025] figure 1 [0026] figure 2 [0027] image 3 Detailed ways [0028] [0029] 2 HPO 4 2 [0030] [0031] [0032] [0033] [0034] [0035] [0036] [0037] [0038] [0039] 10 [0040] [0041] [0042] [0043] [0044] [0045] [0046] [0047] [0048] [0049] [0050] [0051] [0052] 2 CO 3The pH value of the solution to 8.0; [0053] 2) Add 500 μL of anti -air intestinal curved bacon monoclonal antibody and 500 μL of Apu Pujueine A solution to the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569
CPCG01N33/54326G01N33/56911G01N33/56916G01N33/56922
Inventor 薛峰滕军蒋原陈伟李东明曾德新夏晓莉邵景东
Owner ANIMAL AND PLANT & FOOD DETECTION CENTER JIANGSU ENTRY EXIT INSPECTION AND QUARANTINE BUREAU
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