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Method for detecting trace ochratoxin A (OTA) by adopting electrochemical aptamer sensor

An aptamer sensor, ochratoxin technology, applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problem that the change of electrical signal is not obvious, and achieve the effect of low detection cost, simple and flexible operation, and small reagent volume

Inactive Publication Date: 2013-12-04
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the low concentration of OTA in the actual sample, the change of the generated electrical signal is not very obvious, so the technical difficulty of the aptamer electrochemical sensor is to find an efficient signal amplification method to realize the sensitive detection of trace OTA.

Method used

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  • Method for detecting trace ochratoxin A (OTA) by adopting electrochemical aptamer sensor
  • Method for detecting trace ochratoxin A (OTA) by adopting electrochemical aptamer sensor
  • Method for detecting trace ochratoxin A (OTA) by adopting electrochemical aptamer sensor

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1: Preparation of aptasensor

[0030] (1) Preparation of DNA3-GNPs: Take 100 mL of 0.01% HAuCl 4 Heat to boiling under stirring, then add 10 mL of 1% sodium citrate, continue heating and stirring for half an hour, then stop heating, stir and cool to room temperature to obtain gold colloid, the transmission electron microscope image of which is shown in figure 1 middle. Take 2 mL of gold gel and add 30 μL of 100 μM DNA3 solution to incubate for 12 h, then add 0.2 mL of 2 M NaCl solution and continue to incubate for 24 h. After the reaction was completed, the obtained solution was washed twice with water, centrifuged at 12000 r / min for 10 min, and repeated three times to obtain DNA3-GNPs. Finally redisperse in 600 μL Tris-HCl (pH 8.0) for later use.

[0031] (2) Gold electrode pretreatment: First, the gold electrode was polished with 1 μm, 0.5 μm, and 0.05 μm alumina powder in sequence, and then treated with K 3 Fe(CN) 6 As a probe scanning peak potential ...

Embodiment 2

[0033] Example 2: Drawing of the standard curve of the aptamer sensor to the trace OTA standard sample

[0034] The aptasensors were first placed in OTA standard solutions with concentrations of 0, 0.001, 0.01, 0.1, 0.5, 1, 2, and 4 ng / mL to react for 15 min, and then rinsed with secondary water for 3 times. Then soak in 80 μM MB solution for 10 min. Rinse with water twice, then use the aptamer sensor as the working electrode, the platinum wire as the counter electrode, and the saturated calomel electrode as the reference electrode. In 50 mM Tris-HCl, use DPV in the potential range of 0~-0.4 V Collect the corresponding MB reduction peak current signal, draw a standard curve, and the results are shown in image 3 middle.

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Abstract

The invention relates to a method for detecting trace ochratoxin A by adopting an electrochemical aptamer sensor, and belongs to the technical field of electrochemical sensors. The method comprises the following steps: firstly, the DNA modification is performed on a gold electrode; then an aptamer is assembled through the complementary base pairing effect so as to capture labeled DNA of gold nanoparticles; the DNA rich in G is assembled on the surfaces of the gold nanoparticles so as to capture a great quantity of methylene blue (MB) to build a signal amplification strategy; finally, a series of OTA standard solutions is detected by taking differential pulse voltammetry scanning as the detection means so as to obtain the response relation between the reductive peak current of MB and the concentration of OTA, and a standard curve is drawn. The invention aims to provide a preparation method of an aptamer sensor, which is simple in operation, high in sensitivity and strong in specificity and is used for sensitivity detection of trace OTA.

Description

technical field [0001] The present invention relates to a method for the preparation of an electrochemical sensor for the detection of trace amounts of ochratoxin A (OTA), characterized by the application of ochratoxin A aptamers and the signal based on gold nanoparticles and base G-rich DNA The construction of an amplification strategy, and the high specificity, high sensitivity and ease of operation of the detection method belong to the technical field of electrochemical sensors. Background technique [0002] Ochratoxin A (OTA) is a toxic metabolite produced by fungi of the genus Ochratoxin, which is a typical food-borne mycotoxin. Pure OTA is a stable colorless crystalline compound with the chemical formula C 20 h 18 ClNO 6 , Slightly soluble in water, easily soluble in dilute sodium bicarbonate solution. OTA can exist stably in polar organic solvents. For example, the ethanol solution of OTA can exist stably for more than one year under refrigerated conditions, but O...

Claims

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Application Information

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IPC IPC(8): G01N27/327
Inventor 王坤钱静杨兴旺
Owner JIANGSU UNIV
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