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Preparation method of aptamer sensor for detecting residual kanamycin in milk

An aptamer sensor and kanamycin technology, applied in the field of biosensors, can solve the problems of long turnaround time, expensive equipment, cumbersome sample pretreatment, etc.

Inactive Publication Date: 2018-11-06
SHANDONG UNIV OF TECH
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Problems solved by technology

[0003] Recently, many analytical methods have been used to detect KAN due to their sensitivity, such as gas chromatography, capillary electrophoresis (CE), high performance liquid chromatography (HPLC), solid phase extraction (SPE), immunoassay, enzyme (ELISA) and colloidal gold test paper However, due to long turnaround time, expensive equipment, cumbersome sample pretreatment and other reasons, the application of most of the above methods is still partially limited, and it is still necessary to select new methods for sensitive and specific analysis and detection of KAN

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  • Preparation method of aptamer sensor for detecting residual kanamycin in milk
  • Preparation method of aptamer sensor for detecting residual kanamycin in milk
  • Preparation method of aptamer sensor for detecting residual kanamycin in milk

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[0025] Example: such as figure 2 As shown, the morphology of OMC-CS and SA-AuNPs composites were characterized by SEM and TEM, respectively, figure 2 (a) shows that OMC-CS is composed of a large number of rod-like particles uniformly and orderly distributed, which makes OMC-CS have high porosity and high surface area, providing a better protective microenvironment for aptamers; SEM image of SA-AuNPs ( figure 2 (b)) showing the separation of AuNPs particles from each other in the presence of streptavidin molecules; as figure 2 As shown in (c), AuNPs particles were successfully labeled with streptomycin; when biotin was added, AuNPs particles aggregated under the high-affinity interaction between streptavidin and biotin groups ( figure 2 (d)).

[0026] Example: use CV to characterize the construction process of the aptasensor ( image 3 ), due to the probe ([Fe(CN) 6 ] 4- / 3- ), the CV of bare GCE shows a pair of well-symmetrical redox peaks (curve a); due to the high ...

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Abstract

An ultra-sensitive electrochemical sensor, based on ordered mesoporous carbon-chitosan (OMC-CS) and gold nano particles-streptavidin (AuNPs-SA), is used for detecting kanamycin (KAN). The AuNPs-SA immobilizes a DNA chain (biotin labeling) by means of the biotin-streptavidin interaction; when being combined with KAN, a DNA 2 chain containing a KAN aptamer is labeled by ferrocene so as to enhance current signal on the surface of an electrode. Some factors that influence the performance of the aptamer sensor are optimized, so that the aptamer sensor has a wide linearity in the range from 1*10<-10> to 4*10<-6> M; detection limit of the KAN can be lowered to 47.2 pM. The aptamer sensor has electrochemical performances such as excellent stability, sensitivity and repeatability. Without interference of competitive analogues, the aptamer sensor shows excellent specificity; besides, the aptamer sensor has been successfully applied to detection of KAN in practical milk samples. The method is hopeful to have more potential to detect more other types of antibiotics.

Description

technical field [0001] The invention provides a method for preparing an aptamer sensor for detecting kanamycin residue in milk, which belongs to the technical field of biosensors. Background technique [0002] Kanamycin (KAN) is an antibiotic purified from Streptomyces. It is widely used in the treatment of Gram-positive and Gram-negative bacteria in humans and animals. Residues of KAN in food can cause severe nephrotoxicity. , ototoxicity and side effects of antibiotic resistance, in order to ensure food safety and quality, the EU has determined the maximum residue limit of KAN is 150μg / kg (257.4 nM), therefore, for the detection of KAN in milk is necessary to ensure our health . [0003] Recently, many analytical methods have been used to detect KAN due to their sensitivity, such as gas chromatography, capillary electrophoresis (CE), high performance liquid chromatography (HPLC), solid phase extraction (SPE), immunoassay, enzyme (ELISA) and colloidal gold test paper Howe...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/30
Inventor 王相友李发兰孙霞郭业民赵文苹
Owner SHANDONG UNIV OF TECH
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