CN108709923B - Electrochemical kit for rapidly visualizing in-vitro disease detection - Google Patents
Electrochemical kit for rapidly visualizing in-vitro disease detection Download PDFInfo
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- CN108709923B CN108709923B CN201810485268.7A CN201810485268A CN108709923B CN 108709923 B CN108709923 B CN 108709923B CN 201810485268 A CN201810485268 A CN 201810485268A CN 108709923 B CN108709923 B CN 108709923B
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
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- G01N27/333—Ion-selective electrodes or membranes
- G01N27/3335—Ion-selective electrodes or membranes the membrane containing at least one organic component
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- G—PHYSICS
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
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Abstract
According to the electrochemical method for rapidly visualizing in-vitro disease detection, after voltage is applied to the two ends of the kit, the probability of the color change rate of a measuring device is an initial color change rate; after fading, adding a sample solution to be tested at the anode, combining specific antigen, antibody and conductive ions in the sample, electrifying again, and then comparing the conductive ions combined with cells with the initial color change rate by measuring the color change rate to obtain qualitative and quantitative detection results, wherein the detection kit can be repeatedly used, has lower cost and simple operation process, can realize quick visual in-vitro detection, and has better application prospect in the fields of food safety, household detection and the like.
Description
Technical Field
The invention belongs to the application of electrochemistry in the field of in vitro detection.
Background
At present, in-vitro rapid diagnosis is widely applied to the fields of medical diagnosis, food detection, pesticide residue, environmental protection judgment and the like in the existing biomarker technology. Such as: gold particle material markers have been widely used in various biological diagnoses, and have the advantages of simple and rapid visual qualitative identification, and low quantitative determination accuracy. The fluorescence diagnostic technique can be used for qualitative and quantitative analysis, but the ultraviolet fluorescence background noise is more, the resolution error is larger, and the quantitative error is larger. Although the up-conversion fluorescence technology solves the problem of noiseless background, can be used for qualitative and quantitative analysis, and is widely used for biological diagnosis of a chromatography kit, the qualitative analysis completely depends on a complex and expensive special detection instrument, the use efficiency is reduced, and the purpose of timely visualization cannot be achieved. The combination of the up-conversion luminescent material, the magnetic material, the nano-silver material, the organic fluorescent dye and the like also requires auxiliary equipment to realize detection, and cannot be effectively applied to the technical field of chromatography detection.
Along with the gradual increase of the application of the nanometer technology, the integration of the microelectronic technology and the biological technology and the novel electrochemical detection method in recent years, a similar patent CN101093220A discloses an in-vitro detection method, a kit and an instrument based on magnetic particles and microelectrodes; fixing a specific marker 1 on magnetic particles, hybridizing a substance to be detected, a specific marker 2 and gold particles in a liquid phase to form a magnetic-gold particle complex and a silver-dyed complex, separating the magnetic-gold particle complex and the silver-dyed complex, transferring the magnetic-gold particle complex and the silver-dyed complex onto a micro electrode, drying the magnetic-gold particle complex and the silver-dyed complex, measuring the surface impedance of the magnetic-gold particle complex by using an electrical detector, and qualitatively or quantitatively analyzing the substance to be detected, and developing an integrated automatic hybridization, silver-dyed and detection instrument and a related kit based on the method and an automatic technology; the method can obtain qualitative and quantitative detection results, but the detection process is complex and rapid detection cannot be realized. Patent CN1928542B discloses an electrochemical method for in vitro detection of epirubicin concentration by using a nanogold electrode; relates to a nano gold electrode and application thereof. Provides a preparation method of a nano-gold electrode with low cost, high sensitivity, good anti-interference performance and high stability and an electrochemical method for using the nano-gold electrode to detect the concentration of epirubicin in vitro. The preparation method comprises the following steps of polishing a bare glassy carbon electrode into a mirror surface after polishing, cleaning and drying the mirror surface, putting the mirror surface into KAuCl4 and KCl electrolyte, and performing cyclic voltammetry scanning to obtain a nanogold electrode which can be used for in-vitro detection of epirubicin concentration, wherein the steps are as follows: adding PBS into an electrolytic cell, adding a epirubicin standard sample solution, taking a nanogold electrode as a working electrode, taking a saturated calomel electrode as a reference electrode, taking a platinum electrode as a counter electrode to form a three-electrode system, scanning and recording the peak current value of a corresponding differential pulse voltammogram, correlating the peak current value with the concentration of the epirubicin standard sample solution to obtain a linear model, and calculating to obtain the concentration of the epirubicin. The detection of the method is rapid and stable, but the cost for detecting the nano-gold electrode is high, so that the method is difficult to popularize on the market.
According to the electrochemical method for rapidly visualizing in-vitro disease detection, after voltage is applied to the two ends of the kit, the probability of the color change rate of a measuring device is an initial color change rate; after fading, adding a sample solution to be tested into the anode, combining specific antigen, antibody and conductive ions in the sample, and applying voltage to two ends of the kit, wherein the transmittance of the kit through a measuring device is an initial color changing rate L0; after the color is faded, a sample solution to be tested is added into the anode, specific antigen, antibody and conductive ions in the sample are combined, the conductive ions combined with the antigen and the antibody cannot pass through the electrochromic film after being electrified again, then the transmittance of the device is measured to be L1, and a qualitative and quantitative detection result v is obtained through the value of the color change rate delta L according to the change delta L = (L0-L1)/L0 of the transmittance.
The invention has the advantages of high detection efficiency, qualitative and quantitative detection results, reusable detection kit, low cost, simple operation process, capability of realizing rapid visual in-vitro detection, and good application prospect in the fields of food safety, household detection and the like.
Disclosure of Invention
An electrochemical method for rapid visualization of in vitro disease detection; the detection kit comprises two layers of electrodes with electrochromic structures, wherein the two layers of electrodes are added at two ends of the detection kit, the electrode connected with a positive electrode is defined as an anode, and the electrode connected with a negative electrode is defined as a cathode; the surface-treated conductive ion solution places the treated conductive ions in the anode area and places the treated electrochromic material in the cathode area; the electrochromic film is treated by an ionic membrane, and the treated electrolyte membrane is arranged between two layers of electrodes; after the electrochromic kit is electrified and detected, the color change of the kit is measured by a CCD system under the irradiation of a light source, and the color change rate is defined as a color change rate delta L, wherein the color change rate probability of a measuring device after voltage is applied to two ends of the kit is an initial color change rate L0; after the color is faded, a sample solution which is subjected to electrochemical treatment and needs to be tested is added into an anode, specific antigen, antibody and conductive ions in the sample are combined, the conductive ions which are combined with the antigen and the antibody cannot pass through the electrochromic film after being electrified again, the conductive ions which are not combined with the antigen and the antibody normally pass through the electrochromic film, then the transmittance of the device is measured to be L1, and qualitative and quantitative detection result ν is obtained according to the value of the color change Δ L according to the change Δ L = (L0-L1)/L0.
Two layers of electrodes added at two ends of the detection kit are respectively connected with the positive electrode and the negative electrode of a direct current power supply, the conductive base materials of the electrodes at two ends are selected from one or more of an ITO transparent conductive film, a graphene transparent conductive film or a nano silver wire transparent conductive film, then the result of in-vitro detection is realized by adding voltage between the two electrodes, and the selected transparent electrode has no influence on the color changing efficiency of the detection result. The surface treated conductive ion solution is prepared by dissolving salt containing conductive ions lithium and sodium into an aqueous solution to prepare a conductive ion solution with the concentration of 0.9 percent, and then forming a biocompatible group on the surface of the conductive ion by surface treatment, wherein the biocompatible group can be mutually connected with a target antigen and an antibody; the movement of the conductive ions can be controlled after the power is supplied.
The electrochromic film is prepared by uniformly mixing PVB (polyvinyl butyral) serving as a carrier material with the carrier material and then carrying out hot pressing on the mixture, and then the surface of the electrochromic film is in accordance with an ion film, only ions are allowed to pass through but substances such as antigens, antibodies and the like cannot pass through the ion film; the PVB carrier is prepared into carrier slurry by taking propylene carbonate as a solvent, the electrochromic material is one or more of PEDOT, Prussian blue and nano tungsten oxide, and then the electrochromic material is added into the carrier slurry according to a certain proportion and is prepared into the electrochromic film in a hot pressing mode.
Drawings
FIG. 1 is a schematic diagram of an electrochemical detection kit,
fig. 2. an electrochromic film,
description of the drawings:
1 anode, a,
2 cathode, a,
3 an electrochromic film,
4 conductive ions,
5 an electrochromic material,
6 a sample adding port,
7 ion membrane.
Detailed Description
An electrochemical method for rapid visualization of in vitro disease detection; the detection kit comprises two layers of electrodes with electrochromic structures, wherein the two layers of electrodes are added at two ends of the detection kit, the electrode connected with a positive electrode is defined as an anode, and the electrode connected with a negative electrode is defined as a cathode; the surface treated conductive ion solution is used for placing the treated conductive ions in an anode region, the ionic membrane treated electrochromic film is used for placing the treated electrolyte membrane between two layers of electrodes, and the treated electrochromic material is placed in a cathode region; after the electrochromic kit is electrified and detected, the color change of the kit is measured by a CCD system under the irradiation of a light source, and the color change rate is defined as a color change rate delta L, wherein the color change rate probability of a measuring device after voltage is applied to two ends of the kit is an initial color change rate L0; after the color is faded, a sample solution which is subjected to electrochemical treatment and needs to be tested is added into an anode, specific antigen, antibody and conductive ions in the sample are combined, the conductive ions which are combined with the antigen and the antibody cannot pass through the electrochromic film after being electrified again, the conductive ions which are not combined with the antigen and the antibody normally pass through the electrochromic film, then the transmittance of the device is measured to be L1, and qualitative and quantitative detection result ν is obtained according to the value of the color change Δ L according to the change Δ L = (L0-L1)/L0.
The two layers of electrodes are added at two ends of the detection kit, conductive base materials of the two ends of the electrodes are selected from one or more of ITO transparent conductive films, graphene transparent conductive films or nano silver wire transparent conductive films, the conductive base materials are respectively connected with the positive electrode and the negative electrode of a direct current power supply, the conductive base materials are connected with the positive electrode and the negative electrode of the direct current power supply respectively, the positive electrode is connected with the positive electrode and is an anode area, the negative electrode is connected with the negative electrode and is a cathode area, electric charges are given out by the anode to drive conductive ions to move towards the cathode, after the conductive ions are contacted with a color-changing material in the cathode area, the electric charges are transferred to cause the color of the color-changing material to change.
The surface-treated conductive ion solution is prepared by dissolving a biological electrolyte salt containing conductive ions in water by taking one or more of sodium chloride, potassium chloride and lithium chloride as raw materials to prepare a 0.9% conductive ion solution, and cells in the aqueous solution with the concentration can keep activity without inactivation, so that the accuracy of a detection result is improved; then, forming a biocompatible group on the surface of the conductive ion through surface treatment, and connecting the biocompatible group with a target antigen and an antibody; the movement of the conductive ions can be controlled after the power is supplied.
The electrochromic film is prepared by uniformly mixing PVB (polyvinyl butyral) serving as a carrier material and then carrying out hot pressing on the mixture to prepare the electrochromic film, then respectively compounding a layer of cation selective permeation film on the surface of the electrochromic film, and only allowing cations to pass through an ionic film under the action of a direct current power supply to generate an electrochromic phenomenon after the kit is electrified, so that the electrochromic material in a cathode region is prevented from reaching an anode through the ionic film; the PVB carrier is prepared into carrier slurry by taking propylene carbonate as a solvent, the electrochromic material is one or more of PEDOT, Prussian blue and nano tungsten oxide, and then the electrochromic material is added into the carrier slurry according to a certain proportion and is prepared into the electrochromic film in a hot pressing mode.
An electrochemical method for rapid visualization of in vitro disease detection; firstly, adding 1-3V direct current voltage at two ends of a kit, defining the color change rate of a cathode area as an initial color change rate by measuring the color change rate, and then adding reverse voltage until the color is changed to an initial state; and then adding the electrochemically treated antigen and antibody test sample into an anode area, fully mixing for 3-5min, applying a voltage, wherein electrolyte ions combined with the antigen and the antibody cannot pass through the ionic membrane due to the existence of the ionic membrane, ions not combined with the antigen and the antibody can reach a cathode area through the ionic membrane to generate a color change reaction, defining the color change rate at the moment as a sample color change rate L1, comparing with an initial color change rate L0, and obtaining a qualitative and quantitative detection result v according to the value of the color change rate delta L according to the change delta L = (L0-L1)/L0 of the transmittance.
An electrochemical method for rapid visualization of in vitro disease detection; can realize rapid and accurate visual detection in the field of in vitro disease detection, and has better application prospect in the fields of food safety, heavy metal ion standard exceeding and the like
Advantages of the invention
Compared with the prior art, the invention has simple preparation process, and the core device structure is prepared by simple hot-pressing film forming, so the cost investment is low.
The operation process of the process is higher in controllability, the whole detection kit can be prepared directly by a microelectronic packaging process, and the industrial continuous production is easy to realize.
The invention has simple process environment requirement, and can be completed only under vacuum condition in the packaging and curing process, thereby having low production cost and high process controllability.
The electrolyte ions adopt a curing film-forming process, so that the use safety performance is greatly improved, the reuse efficiency is improved, and the use cost is reduced.
Claims (2)
1. An electrochemical kit for rapidly visualizing in-vitro disease detection is characterized by comprising an electrode formed by two layers of transparent conductive films, a conductive ion solution subjected to surface treatment and an electrochromic film subjected to ion film treatment; conducting ion solution containing lithium or sodium is subjected to surface treatment to form a biocompatible group which can be mutually connected with target cells or viruses, and the movement of conducting ions can be controlled after electrification; the treated electrochromic film is placed between two layers of transparent conductive films, an electrode connected with a positive electrode is defined as an anode, and an electrode connected with a negative electrode is defined as a cathode; placing the treated conductive ion solution in an anode region, and placing the treated electrochromic film in a cathode region to form an electrochromic kit; measuring luminous flux of the discolored kit by using an optical processing system under the irradiation of a light source to define the discoloring rate delta L and the disease infection rate v; when the kit is used, a sample to be detected is added into an anode region, voltage is applied to two ends of the kit, and the transmittance of a measuring device is an initial color change rate L0; and adding a sample solution to be tested into the anode after the color is faded, combining specific target cells or viruses in the sample with conductive ions, measuring the transmittance of the device as L1 when the conductive ions combined with the target cells or viruses cannot pass through the electrochromic film after the sample solution is electrified again, and obtaining a qualitative and quantitative detection result v according to the change of the transmittance, namely the color change rate delta L = (L0-L1)/L0.
2. The electrochemical kit for rapid visualization of in vitro disease detection according to claim 1, wherein the electrochromic film treated with ionic membrane is prepared by using PVB as carrier slurry and propylene carbonate as solvent, then adding electrochromic material into the carrier slurry according to a certain proportion and preparing the electrochromic film by hot pressing, and each ionic membrane is covered on the surface of the electrochromic film, so that only ions are allowed to pass through but cells or viruses cannot pass through; the electrochromic material is one or more of PEDOT, Prussian blue and nano tungsten oxide.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH01291221A (en) * | 1988-05-19 | 1989-11-22 | Canon Inc | Electrochromic element |
| CN1790146A (en) * | 2005-11-24 | 2006-06-21 | 同济大学 | Electrochromism device with polyaniline-polyvinyl alcohol compound film and preparation method thereof |
| CN101514958A (en) * | 2008-02-19 | 2009-08-26 | 佳世达科技股份有限公司 | Fluid measuring device |
| CN105218864A (en) * | 2015-09-07 | 2016-01-06 | 四川大学 | A kind of Electrochromic composite material based on nano-cellulose and device preparation method |
| CN105838350A (en) * | 2016-04-16 | 2016-08-10 | 吉林大学 | Electrochromic composite material and prepared electrochromic device |
| CN106093148A (en) * | 2016-06-06 | 2016-11-09 | 济南大学 | A kind of visualization light sends a telegraph H in biosensor detection cell2the construction method of S |
| CN207037308U (en) * | 2017-06-12 | 2018-02-23 | 上海洞舟实业有限公司 | A kind of flexible solid-state electrochromic device |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2746934B1 (en) * | 1996-03-27 | 1998-05-07 | Saint Gobain Vitrage | ELECTROCHEMICAL DEVICE |
| CN102300501B (en) * | 2009-02-04 | 2015-04-08 | 艾伯特糖尿病护理公司 | Multi-function analyte test device and methods therefor |
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Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH01291221A (en) * | 1988-05-19 | 1989-11-22 | Canon Inc | Electrochromic element |
| CN1790146A (en) * | 2005-11-24 | 2006-06-21 | 同济大学 | Electrochromism device with polyaniline-polyvinyl alcohol compound film and preparation method thereof |
| CN101514958A (en) * | 2008-02-19 | 2009-08-26 | 佳世达科技股份有限公司 | Fluid measuring device |
| CN105218864A (en) * | 2015-09-07 | 2016-01-06 | 四川大学 | A kind of Electrochromic composite material based on nano-cellulose and device preparation method |
| CN105838350A (en) * | 2016-04-16 | 2016-08-10 | 吉林大学 | Electrochromic composite material and prepared electrochromic device |
| CN106093148A (en) * | 2016-06-06 | 2016-11-09 | 济南大学 | A kind of visualization light sends a telegraph H in biosensor detection cell2the construction method of S |
| CN207037308U (en) * | 2017-06-12 | 2018-02-23 | 上海洞舟实业有限公司 | A kind of flexible solid-state electrochromic device |
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