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Method for high-sensitively detecting microcystin by employing dynamic light scattering

A microcystin and dynamic light scattering technology, applied in the field of immunoassay chemistry, can solve the problems of poor specificity, no commercial supply of specific protein phosphatase, unable to meet the requirements of rapidity, convenience and accuracy, etc. The effect of simplifying the conditions of the reaction

Inactive Publication Date: 2009-03-25
JIANGNAN UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

Instrumental analysis methods mainly include high-performance liquid chromatography (HPLC), mass spectrometry, liquid chromatography-mass spectrometry, etc. Although these methods are sensitive, they require expensive instruments and equipment, professional operators, and relatively high requirements for inspection materials. Further sample pretreatment can be carried out, which can no longer meet the requirements of modern detection for fast, convenient and accurate
The biochemical method is mainly the protein phosphatase inhibition test method, which has the advantage of being fast and can detect a large number of samples in a few hours, but its biggest disadvantage is that specific protein phosphatases are not commercially available and have poor specificity

Method used

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  • Method for high-sensitively detecting microcystin by employing dynamic light scattering
  • Method for high-sensitively detecting microcystin by employing dynamic light scattering
  • Method for high-sensitively detecting microcystin by employing dynamic light scattering

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Experimental program
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Effect test

Embodiment 1

[0042] (1) Preparation of immunogen:

[0043]① Take 0.25 mL of ethanol solution dissolved with microcystin-LR (MC-LR), add 0.75 mL of deionized water, and add 150 μL of carbodiimide (EDC) to obtain liquid A.

[0044] ② Dissolve 2 mg of bovine serum albumin (BSA) in 1 mL of 25% ethanol solution to obtain liquid B.

[0045] ③ Add liquid A to liquid B dropwise, then add 150 μL of carbodiimide (EDC), mix well, and react at 4°C for 12 hours.

[0046] (2) Preparation of the original coating:

[0047] ① Dissolve 0.1mg of MC-LR in 0.1mL of N,N-dimethylformamide (DMF), cool to 4°C, add 4.8μL of tributylamine, and mix well.

[0048] ② Add 2.7 μL of isobutyl chloroformate to the above solution, mix well, stir and react at 4°C for 20 min to obtain liquid A.

[0049] ③ Dissolve 0.5 mg of ovalbumin (OVA) in 900 μL of borate buffer solution with pH=9.0 and 50 mmol / L, cool to 4°C, and mix well to obtain solution B.

[0050] ④ Slowly add solution A to solution B dropwise, and react at 4°C ...

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Abstract

The invention discloses a method for detecting microcystin-LR in a high sensitivity mode by applying dynamic light scatting, which belongs to the technical field of immunoassay chemistry. A gold nanoparticle synthesized by the method couples with a coating antigen of microcystin to obtain a gold ligated coating antigen, and is connected with an antibody against the microcystin to obtain a gold ligated antibody, the antibody and the gold nanoparticle are used for an immune reaction, trisodium citrate is utilized to reduce a dimmer, an oligomer, and a coagulation formed by the reaction of the gold ligated antibody, the gold ligated coating antigen and the microcystin in a liquid environment, and the content of the microcystin is measured by measuring particle diameter distribution in the liquid environment through dynamic light scattering. The method reacts in liquid environments only, does not need the step of washing, only needs a one-step reaction, simplifies reaction conditions, improves the sensitivity of the detection, and reaches the leading level in the world.

Description

technical field [0001] The invention discloses a method for highly sensitive detection of microcystin by dynamic light scattering, which belongs to the technical field of immunoanalysis chemistry. Background technique [0002] With the development of the economy, sewage containing a large amount of nitrogen and phosphorus nutrients enters lakes and reservoirs, causing eutrophication of water bodies, causing abnormal proliferation of algae, releasing secondary metabolites algae toxins (Algae Toxins), threatening the safety of drinking water for humans and the safety of other organisms in the water. Among them, the wide distribution and great harm of Microcystins (MCs) should be paid more attention. It is a structurally similar cyclic heptapeptide family produced by certain species of freshwater cyanobacteria, mainly Microcystis aeruginosa, and more than 60 isomers are known. Its structure is as follows: [0003] [0004] It contains five fixed amino acids and two variab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/47
Inventor 胥传来徐丽广马伟李灼坤
Owner JIANGNAN UNIV
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