Method for rapidly detecting malachite green in aquatic products by Aptamer

A malachite green and aptamer technology, applied in the direction of material inspection products, biochemical equipment and methods, biological testing, etc., can solve the problems of aquaculture farmers' weak awareness of healthy breeding, imperfect aquatic product safety management mechanism, monoclonal It is difficult to obtain antibodies and other problems, and it is easy to promote and apply on a large scale, conducive to large-scale production, and easy to refold.

Inactive Publication Date: 2009-02-04
CHINA JILIANG UNIV +1
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Problems solved by technology

[0003] In view of the harmfulness of malachite green and its metabolites, many countries have listed it as a banned drug for aquaculture, and my country also listed it in the "List of Veterinary Drugs and Their Compounds Banned for Food Animals" in May 2002. However, Due to the weak awareness of aquaculture farmers' healthy farming and the imperfect safety management mechanism of aquatic products, in 2005 alone, various aquatic products (such as: eel, eel, High residues of malachite green were detected in soft-shelled turtles, etc.
[0004] At present, there are mainly three kinds of detection methods about malachite green and its metabolites, but these m

Method used

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  • Method for rapidly detecting malachite green in aquatic products by Aptamer
  • Method for rapidly detecting malachite green in aquatic products by Aptamer
  • Method for rapidly detecting malachite green in aquatic products by Aptamer

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Embodiment Construction

[0034] Preparation of malachite green aptamer

[0035] (1) Construct a random ssDNA library with a sequence length of about 80 (range: 75 to 90) bases, with fixed sequences with restriction endonuclease sites at both ends of the ssDNA sequence, and the ssDNA sequence is: ( 5'ATAGAGCTCATGGAGTCTCCCTCGG-(N35)-TTTGGATCCGGCAGTGGTGGGCGGGC3'), with 35 random bases (N35) in the middle, and PCR primers were designed according to the fixed sequences at both ends of the ssDNA sequence;

[0036] Upstream primer: 5'-ATAGAGCTCATGGAGTCTCC-3',

[0037] Downstream primer: 5'-GCCCGCCCACCACTGCCGG-3'

[0038] (2) The asymmetric PCR method is used to amplify the ssDNA library obtained in step (1).

[0039] The operation process of asymmetric PCR method is as follows:

[0040] Introduce different concentrations of primers in the PCR amplification cycle (generally, the primer concentration in the ratio of 50:1-100:1 is used), and the main product is double-stranded DNA (dsDNA) in the first 10-15 cy...

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Abstract

The present invention discloses a method which adopts the aptamer rapidly to detect the malachite green in water products. The sample to be detected, the malachite green aptamer and the report sequence with detection markers are mixed; after the report sequence and the malachite green are competitively combined with the malachite green aptamer, the markers on the report sequence is detected, and the content of the malachite green in the sample to be detected is calculated. The kit and the fast detecting test paper, which are prepared according to the method, can better satisfy the current urgent needs for detecting the malachite green residue in the water products, can be used in the detection of the customs and the food sanitation department, and can be easily promoted and applied in a large scale.

Description

technical field [0001] The invention relates to the field of food detection, in particular to a method for detecting malachite green in aquatic products. Background technique [0002] Malachite green (Malachite green, referred to as MG), belongs to triphenylmethane dyes, and its metabolite is colorless malachite green (Leucomalachite green, LMG). For a long time, because malachite green has good antifungal properties in aquaculture disease, freshness of aquatic products and low price, etc. Therefore, it has become the "fishing medicine" favored by many farmers to use, but these two substances have side effects such as high toxin, high residue, high teratogenicity and mutagenicity, not only It has brought huge obstacles to the export of aquatic products in our country, and more importantly, it has constituted a potential threat to people's health. [0003] In view of the harmfulness of malachite green and its metabolites, many countries have listed it as a banned drug for aq...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N33/52C12Q1/68
Inventor 张明洲刘军胡华军王墨染方灵黄军
Owner CHINA JILIANG UNIV
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