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Method for detection of ochratoxin A with ultralow concentration by label-free aptamer sensor

An ochratoxin, ultra-low concentration technology, applied in measuring devices, instruments, material analysis by electromagnetic means, etc., can solve the problems of high detection cost, complex detection scheme, low sensitivity, etc., and achieve the effect of reducing cost

Inactive Publication Date: 2014-01-15
JIANGSU UNIV
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Problems solved by technology

[0005]Technical problem: The purpose of this invention is to invent an electrochemical aptasensor that integrates the advantages of label-free, high sensitivity, high selectivity, simple and easy operation, etc. , to provide a method for quantitative detection of OTA with simple preparation process, high sensitivity, wide measurement range and low cost, and solve the existing problems of high detection cost, complicated detection scheme, long detection time and low sensitivity

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  • Method for detection of ochratoxin A with ultralow concentration by label-free aptamer sensor
  • Method for detection of ochratoxin A with ultralow concentration by label-free aptamer sensor
  • Method for detection of ochratoxin A with ultralow concentration by label-free aptamer sensor

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Embodiment 1

[0028] Preparation of reduced graphene oxide composites

[0029] (1) Surface thiolation of graphene oxide: firstly, graphene oxide (GO) was prepared by the Hummers method, and then 25 mg GO was mixed with 0.5 g 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide Hydrochloride (EDC) and 0.5 g N-hydroxysuccinimide (NHS) were mixed in 50 mL ethanol solution and reacted at room temperature for 8 h with constant stirring. Then add 1.0 g of o-aminothiophenol into the above reaction system and continue to stir for 12 h at room temperature. Several times, thiolated graphene oxide (GO-SH) was obtained after vacuum drying.

[0030] (2) Synthesis of gold nanoparticles-modified GO-SH composites: 10 mg GO-SH was ultrasonically dispersed in 20 mL double-distilled water, then transferred to a three-hole flask and heated to reflux for 0.5 h while stirring. Add 5 mL of freshly prepared sodium citrate solution (25 mg mL –1 ) was slowly added into the three-hole flask, and continued to reflux for...

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Abstract

The invention relates to a method for detection of ochratoxin A (OTA) with ultralow concentration by a label-free aptamer sensor. The method comprises the following specific steps: using o-amino thiophenol functionalized graphene oxide (GO-SH) and chloroauric acid as starting materials and sodium citrate as a reducing agent to prepare gold nanoparticles modified functionalized reduction graphene oxide (Au-S-rGO) by a one pot method; loading mercapto modified single stranded DNA3 by using the Au-S-rGO as a vector to obtain DNA3-Au-S-rGO; then modifying mercapto modified single stranded DNA1 on a gold electrode surface, and modifying an aptamer (DNA2) to the electrode surface through complementary pairing of bases; and then reacting the sensor interface with OTA of different concentrations. The method for quantitative detection of OTA has the advantages of simple preparation process, high sensitivity, wide measuring range and low cost, and solves the problems of high cost, complex detection scheme, long detection time and low sensitivity in the prior art.

Description

technical field [0001] The invention relates to a synthesis method of a nano-gold modified mercapto-reduced graphene oxide composite material and the preparation of an electrochemical aptamer sensor used as a marker for ultra-low concentration ochratoxin A (ochratoxin A, OTA) detection The method belongs to the technical fields of biomolecular recognition and nanomaterial-assisted signal amplification. Background technique [0002] OTA is a mycotoxin with multiple toxicity produced by the metabolism of Aspergillus and Penicillium. It has strong stability and is not easy to degrade. It widely exists in grain, feed and cereal food. Animals eating feed containing OTA will lead to residual toxins in the body, which will contaminate meat products. These residual OTAs can also enter the human body through the food chain, thus posing a serious threat to human health and agricultural economic development. Research data show that OTA has nephrotoxicity, hepatotoxicity, immunotoxicit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327
Inventor 王坤钱静姜玲
Owner JIANGSU UNIV
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