A kind of magnetic enrichment material, water body bacteria detection kit and application

A technology of bacteria detection and kits, which is applied in the direction of magnetic materials, magnetic objects, microbial measurement/inspection, etc., and can solve problems such as difficult to meet rapid detection, cumbersome operation, high cost and technical level requirements

Active Publication Date: 2021-08-17
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cost and technical level of the PCR method are high, and the operation is cumbersome, requiring multiple steps such as bacterial lysis, nucleic acid extraction, and amplification.
In addition, although the PCR method has greatly shortened the detection period of bacteria (3.5-10 hours) compared with the traditional bacterial culture method, it is still difficult to meet the actual on-site rapid detection needs.

Method used

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  • A kind of magnetic enrichment material, water body bacteria detection kit and application
  • A kind of magnetic enrichment material, water body bacteria detection kit and application
  • A kind of magnetic enrichment material, water body bacteria detection kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] In this embodiment, a magnetic nano temperature-sensitive enrichment reagent and a preparation method thereof are provided.

[0056] The preparation method of the magnetic nano temperature-sensitive enrichment reagent is as follows:

[0057] Take 62.2mgFe 3 o 4 , add 5ml of ethanol, 4mL of water, sonicate until completely dispersed. After the supernatant was removed by centrifugation, 10 mL of 0.2 M tetramethylammonium hydroxide solution was added, shaken for 5 min, and ultrasonicated until completely dissolved. Add 4ml of isopropanol and centrifuge to remove the supernatant. Wash twice with 6 ml acetone.

[0058] Take 127.0mg of poly(N-isopropylacrylamide) (PNIPAM) and add water to sonicate until completely dissolved, add the treated Fe 3 o 4 , shaking reaction at room temperature for 3h, the polymer-protected Fe 3 o 4 . The product was centrifuged to remove water. Wash twice with 500 µL of acetonitrile. Add 500 μl of solution B, sonicate until completely di...

Embodiment 2

[0062] This embodiment provides a kit for rapid detection of bacterial content in water bodies, specifically a kit for rapid detection of bacterial contamination in water bodies based on nano-gold click reduction. The kit includes:

[0063] (1) magnetic nano temperature-sensitive enrichment reagent described in embodiment 1;

[0064] (2) eluent;

[0065] (3) Color developer.

[0066] Wherein, the eluent is sterilized 10mM PBS buffer (PH=7.4), which also includes 8mM Na 2 HPO 4 , 136mM NaCl, 2mM KH 2 PO 4 , 2.6 mM KCl.

[0067] The color developer includes color developers C1, C2 and copper sulfate; wherein C1 is an alkynyl-modified gold nano-suspension solution, and C2 is an azide-modified gold nano-particle solution.

[0068] In the developer, C1, C2, CuSO 4 The concentration is 200mM.

[0069] The preparation method of developer C1 and C2 is also provided in this embodiment, and the specific steps are as follows:

[0070] Get 100ml 0.006% chloroauric acid (HAuCl 4 ...

Embodiment 3

[0073] In the present embodiment, provide a kind of method that bacteria in water body are carried out rapid detection, described method is detected by the kit in embodiment 2, comprises the following steps:

[0074] (1) Take 5 mL of the water sample to be tested and centrifuge at 5000r for 2 minutes;

[0075] (2) Take 1 mL of the supernatant to resuspend the precipitate, and discard the rest of the supernatant;

[0076] (3) Add magnetic nanometer temperature-sensitive enrichment reagent (100 μL) to the resuspension obtained in step (2), and shake for 30 minutes. The bacteria liquid is in full contact with the magnetic iron ferric oxide nanoparticles, and the bacteria are specifically captured by concanavalin A on the surface of the polymer. Use a magnet to attract the magnetic nanomaterials to the side of the wall, and remove the supernatant with a pipette gun.

[0077] (4) Add 0° C. eluent (100 μL) to the magnetic nanoparticles obtained in step (3), and shake rapidly (2 mi...

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Abstract

The disclosure belongs to the technical field of bacteria detection, and in particular relates to a magnetic enrichment material, a water body bacteria detection kit and applications. The disclosure provides a magnetic temperature-sensitive enrichment material, which uses thermosensitive polymer poly(N-isopropylisoacryloyl) and concanavalin A to modify magnetic ferric iron tetroxide nanoparticles. The affinity of legumin A to bacteria realizes the adsorption of bacteria in the water environment, and the detachment of bacteria is realized through the temperature-raising phase transition of thermosensitive polymers. The present disclosure also provides a kit including the above-mentioned magnetic temperature-sensitive enrichment material, and also includes a reagent for developing color of the desorbed bacteria. The kit can realize the visual detection of bacteria in the water body. If the color solution changes, it proves that there are bacteria, and the speed of color change is directly proportional to the concentration of bacteria. The kit has fast detection speed and high sensitivity, and can be used in water quality monitoring, water pollution prevention and control, post-disaster epidemic prevention and control and other fields.

Description

technical field [0001] The disclosure belongs to the technical field of bacteria detection, and in particular relates to a magnetic temperature-sensitive enrichment material, a water body bacteria detection kit including the enrichment material, and applications in the fields of water body environment monitoring and water pollution prevention and control. Background technique [0002] The information disclosed in this Background section is only intended to increase the understanding of the general background of the disclosure, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes prior art that is already known to those skilled in the art. [0003] Bacteria are an important class of pathogens. After they invade the body, they will secrete a large amount of biological toxins, thereby destroying the structure and function of the body, causing host infection, and causing various diseases such as gastroenteritis, pneumo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): H01F1/00H01F41/00C12Q1/04
CPCC12Q1/04H01F1/0045H01F41/00
Inventor 杨燕美钱玉珍代莹莹赵迪
Owner SHANDONG NORMAL UNIV
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