Method for detecting ochratoxin A by using electrochemical luminous adaptor sensor

An aptamer sensor, ochratoxin technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of high detection cost, high antibody cost, tedious and time-consuming, etc., to achieve improved sensitivity, high sensitivity, The effect of improving sensitivity and stability

Inactive Publication Date: 2011-01-05
JIANGNAN UNIV
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  • Claims
  • Application Information

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Problems solved by technology

However, antibodies are easily affected by external conditions, especially temperature, and require harsh storage conditions, which largely limits the flexible application of the method
In addition, antibody preparation requires animal experiments or cell experiments, which is tedious and time-consuming, the cost of antibody preparation is high, and the detection cost of the developed method is correspondingly high

Method used

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  • Method for detecting ochratoxin A by using electrochemical luminous adaptor sensor
  • Method for detecting ochratoxin A by using electrochemical luminous adaptor sensor
  • Method for detecting ochratoxin A by using electrochemical luminous adaptor sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Establishment of Ochratoxin A Detection Standard Curve and Detection of Wheat Actual Samples

[0029] The prepared electrochemiluminescent sensor was placed in the ochratoxin A standard solution for reaction, and the concentration of the standard solution was 0 ng / mL, 0.02 ng / mL, 0.04 ng / mL, 0.06 ng / mL, 0.08 ng / mL, 0.1ng / mL, 0.2ng / mL, 0.8ng / mL, 1ng / mL, 1.5ng / mL, 2ng / mL, 3ng / mL, the working voltage is 0.8V, and the working buffer system is Na 2 CO 3 -NaHCO 3 (pH 11.0), each concentration was reacted for 30min, and then added H 2 o 2 The concentration is 1.5mM, and a standard curve is established according to the relative electrochemiluminescence value and the corresponding concentration of ochratoxin A standard substance.

[0030] Sample pretreatment: crush and sieve wheat at high speed, weigh 20g into a 100mL flask, add 5g NaCl, 80% ethanol aqueous solution, mix thoroughly, place in a homogenizer for high-speed stirring and extraction for 2min, stand stil...

Embodiment 2

[0036] Example 2: Detection of ochratoxin A in actual corn samples and experiment of standard addition recovery rate

[0037] The establishment of the standard curve for the detection of ochratoxin A and the sample pretreatment are the same as in Example 1.

[0038] Choose 10 kinds of different types of corn from the farmer’s market, first use the method of the present invention to detect the content of ochratoxin A to obtain the background value, and then select 0.5 μg / kg, 1.0 μg / kg, 2.0 μg / kg, 5.0 μg / kg, 10μg / kg, 5 kinds of OTA standard substances with different concentrations were added to the analyte, and the OTA content was detected again by the method of the present invention to obtain the detection value. Recovery %=(detection value-background value) / addition amount×100%. From the results in Table 2, it is shown that the present invention is stable, sensitive and accurate, and is suitable for the detection of ochratoxin A in actual corn samples.

[0039] Table 2: Det...

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Abstract

The invention discloses a method for detecting ochratoxin A by using an electrochemical luminous adaptor sensor, which belongs to the technical field of electrochemical luminous sensors and is used for detecting the ochratoxin A content of wheat, grains, feeds and products thereof. The method is based on the ochratoxin A specificity identification by an adaptor and an electrochemical luminous reaction of hydrogen peroxide (H2O2) and isoluminol (ABEI) under an alkaline condition. The method comprises the following steps of: modifying the surface of a naked gold electrode by using gold nanoparticles to amplify an electrochemical luminous signal; modifying the surface of a working electrode by using single stranded DNA, and modifying the electrode surface with the adaptor marked with the isoluminol (ABEI) by base pairing; adding the hydrogen peroxide (H2O2) to detect the electric luminous signal; and establishing a quantitative detection standard curve of the ochratoxin A according to the ratio of the strength of the electrochemical luminous signal to the concentration of the ochratoxin A. The invention aims to provide a high-sensitivity and high-operability ochratoxin A quantitative detection method.

Description

technical field [0001] The invention relates to the detection of ochratoxin A by using an electrochemiluminescence sensor, and is characterized in the application of ochratoxin A aptamers and isoluminol electrochemiluminescent markers, and the method of amplifying signals of gold nanoparticles in the process of electrochemiluminescence The function, as well as the high sensitivity and easy operation of the detection method, belong to the technical field of electrochemiluminescence sensors. Background technique [0002] Ochratoxin A (ochratoxin A, OTA) is widely distributed in nature, highly toxic, and extremely harmful to humans, animals and plants. OTA mainly exists in cereals, beans and soy products, dried fruits, coffee, grapes and wine, fragrant plants, oil crops, beer, tea, etc. Toxicological studies have shown that ochratoxin A has renal toxicity, liver toxicity, immunotoxicity, teratogenicity, carcinogenicity and mutagenicity, etc., and has great potential harm to an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/403G01N27/48
Inventor 王周平段诺混旭吴世嘉
Owner JIANGNAN UNIV
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