A method for the detection of trace amounts of ochratoxin a by electrochemical aptasensor

An aptamer sensor and ochratoxin technology, which can be applied to measurement devices, scientific instruments, instruments, etc., can solve the problem that electrical signal changes are not very obvious, and achieve the effects of low detection cost, fast analysis speed, and simplified modification process.

Inactive Publication Date: 2015-08-26
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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  • A method for the detection of trace amounts of ochratoxin a by electrochemical aptasensor
  • A method for the detection of trace amounts of ochratoxin a by electrochemical aptasensor
  • A method for the detection of trace amounts of ochratoxin a by electrochemical aptasensor

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Example 1: Preparation of aptamer sensor

[0029] (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 to heat and stir to react for half an hour, then stop heating, stir and cool to room temperature to obtain gold glue, and its transmission electron microscope image 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 12,000 r / min for 10 min, and repeated three times to prepare DNA3-GNPs. Finally re-dispersed in 600 μL Tris-HCl (pH 8.0) for use.

[0030] (2) Gold electrode pretreatment: First, the gold electrode was polished with 1 μm, 0.5 μm, and 0.05 μm alumina powder in turn, and K 3 Fe(CN) 6 As a probe scanning peak potential difference is less tha...

Embodiment 2

[0032] Example 2: Drawing the standard curve of the aptamer sensor to the trace OTA standard sample

[0033] The aptamer sensor was first placed in the OTA standard solution with concentrations of 0, 0.001, 0.01, 0.1, 0.5, 1, 2, and 4 ng / mL, respectively, for 15 min, and then rinsed three times with secondary water. Then soak in 80 μM MB solution for 10 min. After washing with water twice, the aptamer sensor was used as the working electrode, the platinum wire was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode. 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 amounts of ochratoxin A by an electrochemical aptamer sensor, belonging to the technical field of electrochemical sensors. Firstly, the gold electrode is modified with DNA; then aptamers are assembled through complementary base pairing to capture DNA labeled with gold nanoparticles; then G-rich DNA is assembled on the surface of gold nanoparticles to capture a large amount of methylene blue (MB) Build a signal amplification strategy. Finally, a series of OTA standard solutions were measured by differential pulse voltammetry scanning, and the response relationship between the reduction peak current of MB and OTA concentration was obtained, and the standard curve was drawn. The present invention aims to provide a method for preparing an aptamer sensor with simple operation, high sensitivity and strong specificity for sensitive 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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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
Inventor 王坤钱静杨兴旺
Owner JIANGSU UNIV
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