Electrochemical biosensor of cadmium ion nucleic acid aptamer and screen-printed electrode

A screen-printed electrode and nucleic acid aptamer technology, applied in the field of detection, can solve the problems of real-time food monitoring, high detection cost, expensive equipment, etc., and achieve the effects of fast analysis speed, low detection cost, and convenient portability

Active Publication Date: 2019-11-26
广州百博生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the main sources of heavy metal pollution in food include environmental pollution and food processing, and the existing traditional detection methods for heavy metal cadmium mainly include ultraviolet spectrophotometry, atomic fluorescence spectroscopy, graphite furnace atomic absorption method, and inductively coupled plasma spectroscopy. Although the traditional detection method can accurately detect trace heavy metal elements, the equipment is expensive and requires trained personnel to operate. The operation is time-consuming and the detection cost is high. It is impossible to monitor food in real time on-site and online. Chemical methods are low in cost, cheap in price, simple in operation, high in sensitivity, easy to miniaturize, on-site and on-line detection of heavy metals is the future development trend

Method used

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  • Electrochemical biosensor of cadmium ion nucleic acid aptamer and screen-printed electrode
  • Electrochemical biosensor of cadmium ion nucleic acid aptamer and screen-printed electrode
  • Electrochemical biosensor of cadmium ion nucleic acid aptamer and screen-printed electrode

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Embodiment 1

[0032] Embodiment 1: Construction of the screen-printed electrode electrochemical biosensor according to the present invention

[0033] (1) Reagents and instruments

[0034] Cadmium nitrate tetrahydrate (Cd(NO 3 ) 2 4H 2 O) Purchased from Sigma with a purity of 99%, the cadmium ion nucleic acid aptamer described in the article was synthesized by Shanghai Bioengineering Co., Ltd. Na 2 HPO 4 12H 2 O, NaH 2 PO 4 2H 2 O, NaCl, KCl, K 4 [Fe(CN) 6 ]·3H 2 O, K 3 [Fe(CN) 6 ], concentrated sulfuric acid, and nitric acid were of analytical grade and purchased from Guangzhou Chemical Reagent Factory. The experimental water was ultrapure water (resistivity: 18.2 MΩ / cm).

[0035] CHI660C electrochemical workstation (Shanghai Chenhua Instrument Co., Ltd.), screen-printed gold electrode (DROPSENS) uses gold as the working electrode (diameter 4mm) and auxiliary electrode, and silver as the reference electrode.

[0036] The sequence of the cadmium ion nucleic acid aptamer is: C...

Embodiment 2

[0045] Example 2: Detection of cadmium ion standard by aptasensor and drawing of its standard curve

[0046] The aptamer screen-printed electrochemical biosensor was rinsed with ultrapure water to remove unbound aptamers. The screen-printed electrodes used gold as the working electrode and auxiliary electrode, and silver as the reference electrode. , K at 5mmol / L 3 [Fe(CN)] 6 , containing 5mmol / L K 3 [Fe(CN)] 6 Differential pulse voltammetry analysis was carried out in the electrolyte, and the DPV diagram of the aptasensor and its peak current value Ipo were recorded. The electrochemical parameters are: working potential 300mV-700mV, scan rate 100mV / s, rest time 2s;

[0047] Incubate the aptamer electrochemical biosensor with a total of 7 cadmium ion standard solutions with concentrations of 0.1, 0.5, 1, 5, 10, 50, and 100 ng / ml in sequence, react at 25°C for 30 minutes, and rinse with ultrapure water after the reaction. Soak for 5min to remove non-specifically adsorbed c...

Embodiment 3

[0049] Embodiment 3: Actual sample detection

[0050] The aptamer sensor described in the present invention is used for actual sample determination and recovery rate detection. In this embodiment, Cestbon purified water is selected as the sample for standard recovery detection, and 1ng / ml, 5ng / ml, 10ng / ml, 50ng / ml cadmium ion standard for DPV analysis, through the standard curve established in Example 2, the concentration of the corresponding example is calculated as the sample concentration of cadmium ion in Yibao purified water. The measurement results are shown in Table 1. The sample recovery rate measured by the electrochemical method is between 85.3%-107.5%, and the average sample recovery rate is 97.9%.

[0051] Table 1: Determination results of cadmium ion content in samples

[0052] Sample serial number Amount added (ng / ml) Detection amount (ng / ml) Recovery rate(%) RSD(%) 1 1 1.075 107.5 5.11 2 5 4.675 93.5 4.75 3 10 8.532 85....

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Abstract

The invention relates to a cadmium ion aptamer and a related screen-printed electrode electrochemical biosensor. The cadmium ion aptamer has a length of 22-28 bases, and contains the nucleic acid sequence GTGATGACCTGC. The invention also provides a screen-printed electrode electrochemical biosensor containing the cadmium ion aptamer. Based on the specific recognition function of aptamer and cadmium ion, the invention employs DPV as the electrochemical detection technology, establishes a method for detecting cadmium ion with an electrochemical aptamer biosensor, and provides a new method and way for cadmium ion detection. The adapter and the electrochemical biosensor provided by the invention have the advantages of high specificity, high sensitivity, convenient operation, low detection cost, applicability to field detection and the like.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to a cadmium ion nucleic acid aptamer and a related screen printing electrode electrochemical biosensor. Background technique [0002] Cadmium (Cd, Cadmium) is a non-essential heavy metal element for the human body. It is a highly toxic and hazardous environmental pollutant with the characteristics of difficult metabolism and strong enrichment. The International Agency for Research on Cancer (IARC) lists cadmium as a human carcinogen. Cadmium is a common catalyst in industrial production. The arbitrary discharge of cadmium-containing wastewater, waste gas, and waste residue is an important cause of heavy metal cadmium pollution in the environment. The main source of cadmium pollution in food is that crop roots absorb and enrich cadmium from the soil. On the other hand, cadmium is enriched in shellfish and other aquatic products and then passed through the food chain to cause human ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/30G01N27/327
CPCG01N27/30G01N27/3275
Inventor 白卫滨李瑜姚冬生孙建霞冉国敬
Owner 广州百博生物科技有限公司
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