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Artificial Antigen and Its Application for Detecting Oryzae oryzae Acidosis

An artificial antigen and hapten technology, applied in the field of immunochemical analysis, to achieve the effect of enhanced immunogenicity, strong specificity and simple synthesis route

Inactive Publication Date: 2020-12-15
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there is no relevant report on toxin hapten in China

Method used

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  • Artificial Antigen and Its Application for Detecting Oryzae oryzae Acidosis
  • Artificial Antigen and Its Application for Detecting Oryzae oryzae Acidosis
  • Artificial Antigen and Its Application for Detecting Oryzae oryzae Acidosis

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Experimental program
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Effect test

preparation example Construction

[0036] Specifically, the synthetic method of the toxin hapten of the above structure comprises the following steps:

[0037] S1. methylhydrazine reacts with compound 1 to obtain compound 2, which has the structural formula The compound 2 has the structural formula

[0038] In this step, methylhydrazine and compound 1 undergo a substitution reaction, the principle of which is known to those skilled in the art through technical manuals or through conventional experimental methods. The present invention does not specifically limit the specific operation of the reaction. Those skilled in the art Regular adjustments can be made according to actual needs.

[0039] S2. Under acidic conditions, the compound 2 reacts with formaldehyde to obtain compound 3, which has the structural formula

[0040] In this step, compound 2 and formaldehyde undergo a carbonylamide condensation reaction under acidic conditions to form a carbon-nitrogen double bond. The reaction principle of the ca...

Embodiment 1

[0060] Embodiment 1: Synthesis of toxin hapten

[0061] S1. methylhydrazine reacts with compound 1 to obtain compound 2, which has the structural formula The compound 2 has the structural formula

[0062] S2. Under acidic conditions, the compound 2 reacts with formaldehyde to obtain compound 3, which has the structural formula

[0063] S3. In the presence of sodium nitrite and acetic acid, the compound 3 is reacted to obtain compound 4, which has the structural formula

[0064] S4. The compound 4 is hydrolyzed to obtain the toxin hapten.

[0065] Wherein, the synthetic method of the compound 1 is: (1) provide the compound A, the molecular formula of the compound A is NH 2 (CH 2 ) n COOEt; (2) the compound A reacts with trimethylsilyl isocyanate to obtain compound B, which has the structural formula (3) The compound B reacts with malonic acid and acetic anhydride to obtain compound C, which has the structural formula (4) The compound C is reacted with phosphorus...

Embodiment 2

[0066] Embodiment 2: the synthesis of toxin immunization antigen

[0067] Get 20 mg of carrier protein (bovine serum albumin, BSA) and dissolve it in 4 ml of phosphate buffer (pH7.4), add 5 mg of toxin hapten, mix well, add 10 mg of EDC (1-(3-dimethylaminopropyl base)-3-ethylcarbodiimide hydrochloride), react at room temperature (25°C) for 2 hours, dialyze to remove small molecules that are not bound to the carrier protein, and collect the dialyzed protein as the toxin immune antigen.

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PUM

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Abstract

Disclosed are methods for preparing a hapten of toxoflavin and an artificial antigen of toxoflavin, wherein a highly specific and sensitive antibody can be obtained from an animal immunized with the prepared artificial antigen. The antigen and antibody prepared and the treatment of a sample by means of this method can provide a simple and quick technical approach for the highly sensitive detection of toxoflavin.

Description

technical field [0001] The invention relates to the technical field of immunochemical analysis, in particular to an artificial antigen for detecting oryzae acidosis and its application. Background technique [0002] Yeasted rice noodle poisoning is a type of bacterial food poisoning with a very high fatality rate in the vast rural areas of our country. Pseudomonas cocos subspecies fermented rice noodle is the cause of fermented rice noodle food poisoning and deteriorating white fungus poisoning. Studies have proved that the pathogenic substance of this fungus is two toxins produced during its growth and reproduction: bongkrekic acid (bongkrekic acid) acid, BA) and toxoflavin (toxoflavin, TF). The two toxins are small molecular substances of fatty acids, which are highly toxic, heat-resistant, and highly toxic, and cannot be destroyed by general cooking methods. Therefore, contaminated food can easily cause poisoning. The production of zymosic acid is much higher than toxof...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D487/04C07K14/765C07K14/77C07K14/795C07K16/44G01N33/53G01N33/532G01N33/558G01N33/58
CPCC07D487/04C07K14/765C07K14/77C07K14/795C07K16/44C07K19/00G01N33/53G01N33/532G01N33/558G01N33/587
Inventor 柯跃斌李金峰黎诚耀丁晔赵芳肖云军刘博超
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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