CN107764880A - A kind of biosensor thin layer electrode of quick detection miocardial infarction protein marker - Google Patents
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- 238000001514 detection method Methods 0.000 title claims abstract description 13
- 239000012474 protein marker Substances 0.000 title claims abstract description 11
- 206010061216 Infarction Diseases 0.000 title 1
- 230000007574 infarction Effects 0.000 title 1
- 208000010125 myocardial infarction Diseases 0.000 claims abstract description 19
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 10
- 239000002105 nanoparticle Substances 0.000 claims abstract description 8
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 8
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 8
- 238000007747 plating Methods 0.000 claims abstract description 6
- 230000008569 process Effects 0.000 claims abstract description 6
- 239000000126 substance Substances 0.000 claims abstract description 6
- 239000000427 antigen Substances 0.000 claims abstract description 5
- 102000036639 antigens Human genes 0.000 claims abstract description 5
- 108091007433 antigens Proteins 0.000 claims abstract description 5
- 239000000758 substrate Substances 0.000 claims abstract description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 4
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 claims abstract description 4
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 4
- 239000011248 coating agent Substances 0.000 claims abstract description 4
- 238000000576 coating method Methods 0.000 claims abstract description 4
- 150000001875 compounds Chemical class 0.000 claims abstract description 4
- 239000002184 metal Substances 0.000 claims abstract description 4
- 229910052751 metal Inorganic materials 0.000 claims abstract description 4
- 229920000642 polymer Polymers 0.000 claims abstract description 4
- 229910052709 silver Inorganic materials 0.000 claims abstract description 4
- 239000004332 silver Substances 0.000 claims abstract description 4
- 230000035945 sensitivity Effects 0.000 abstract description 9
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 206010028851 Necrosis Diseases 0.000 description 4
- 230000017074 necrotic cell death Effects 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 3
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- 210000004165 myocardium Anatomy 0.000 description 3
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- 210000004351 coronary vessel Anatomy 0.000 description 2
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- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 239000010931 gold Substances 0.000 description 2
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- 238000012360 testing method Methods 0.000 description 2
- 206010003211 Arteriosclerosis coronary artery Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102000004420 Creatine Kinase Human genes 0.000 description 1
- 108010042126 Creatine kinase Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 108010044467 Isoenzymes Proteins 0.000 description 1
- 206010024119 Left ventricular failure Diseases 0.000 description 1
- 102000036675 Myoglobin Human genes 0.000 description 1
- 108010062374 Myoglobin Proteins 0.000 description 1
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- 206010037660 Pyrexia Diseases 0.000 description 1
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- 208000007536 Thrombosis Diseases 0.000 description 1
- 102000013394 Troponin I Human genes 0.000 description 1
- 108010065729 Troponin I Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
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- 238000003556 assay Methods 0.000 description 1
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- G01—MEASURING; TESTING
- 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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- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
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Abstract
一种快速检测心肌梗塞蛋白标记物的生物传感器薄层电极，它包括一块基板，基板上通过化学涂覆或电化学电镀工艺制备有至少一条形薄层电极，在所述薄层电极上制有至少一层含有与心肌梗塞蛋白标记物抗原相互作用、从而能够产生被检测电信号的抗体试剂层；所述抗体试剂层是通过无机凝胶层粘合在薄层电极表面；所述的薄层电极上面还粘合有纳米粒子含有层，并与抗体试剂层共同组成一叠层体；所述的纳米粒子含有层由电化学上为惰性的高分子化合物制成；所述的薄层电极为金属层、碳电极层以及银电极层之一；它具有结构简单、制作方便、成本更低且灵敏度高等特点。
A biosensor thin-layer electrode for rapid detection of myocardial infarction protein markers, which includes a substrate, on which at least one strip-shaped thin-layer electrode is prepared by chemical coating or electrochemical plating process, and the thin-layer electrode is formed on the thin-layer electrode At least one layer contains an antibody reagent layer that interacts with the myocardial infarction protein marker antigen to generate a detected electrical signal; the antibody reagent layer is bonded to the surface of the thin-layer electrode through an inorganic gel layer; the thin layer The electrode is also bonded with a nanoparticle-containing layer, and forms a laminate together with the antibody reagent layer; the nanoparticle-containing layer is made of an electrochemically inert polymer compound; the thin-layer electrode is One of the metal layer, carbon electrode layer and silver electrode layer; it has the characteristics of simple structure, convenient manufacture, lower cost and high sensitivity.
Description
技术领域technical field
本发明涉及的是一种快速检测心肌梗塞蛋白标记物的生物传感器薄层电极，属于生物传感器技术领域。The invention relates to a biosensor thin-layer electrode for rapidly detecting myocardial infarction protein markers, and belongs to the technical field of biosensors.
背景技术Background technique
心肌梗塞是指心肌的缺血性坏死，为在冠状动脉病变的基础上，冠状动脉的血流急剧减少或中断，使相应的心肌出现严重而持久地急性缺血，最终导致心肌的缺血性坏死，现已成为严重影响人类健康的心血管疾病之一。发生急性心肌梗塞的病人，在临床上常有持久的胸骨后剧烈疼痛、发热、白细胞计数增高、血清心肌酶升高以及心电图反映心肌急性损伤、缺血和坏死的一系列特征性演变，并可出现心律失常、休克或心力衰竭。急性心肌梗塞实质上就是心肌的缺血性坏死，其基本原因就是冠状动脉粥样硬化，逐渐发展至管腔狭窄甚至堵塞，在此基础上突然发生血栓，导致冠状动脉完全阻塞所致。急性心律失常、急性左心衰竭极易引起猝死，因此，救治越早，生存机会就越大，早抢救1小时甚至10分钟，都有可能使一个人的生命得到延续。Myocardial infarction refers to ischemic necrosis of the myocardium. On the basis of coronary artery disease, the blood flow of the coronary artery is sharply reduced or interrupted, resulting in severe and persistent acute ischemia of the corresponding myocardium, which eventually leads to myocardial ischemia. Necrosis has become one of the cardiovascular diseases seriously affecting human health. Patients with acute myocardial infarction often have persistent severe pain behind the sternum, fever, increased white blood cell count, elevated serum myocardial enzymes, and electrocardiograms reflect a series of characteristic changes of acute myocardial injury, ischemia, and necrosis. Arrhythmia, shock, or heart failure occurs. Acute myocardial infarction is essentially ischemic necrosis of the myocardium. The basic cause is coronary atherosclerosis, which gradually develops to stenosis or even blockage of the lumen. On this basis, sudden thrombus occurs, resulting in complete blockage of the coronary artery. Acute arrhythmia and acute left heart failure can easily lead to sudden death. Therefore, the earlier the treatment, the greater the chance of survival. Early rescue for 1 hour or even 10 minutes may prolong a person's life.
目前我国心肌梗塞的诊断标准包括：临床诊断、心电图异常及蛋白标志物含量测定三项中的两项；但由于大约25%的心肌梗塞病人发病早期没有典型的临床症状，约50%的病人心电图无典型变化，若单独依靠心电图改变和临床症状，心肌梗塞的诊断符合率仅为75%。蛋白标志物测定具有特异性强、灵敏度高的优点；所以近年来对于心肌梗塞蛋白标志物的研究十分活跃。心肌梗塞的蛋白标志物主要包括肌红蛋白、肌酸激酶同工酶和肌钙蛋白Ⅰ。At present, the diagnostic criteria for myocardial infarction in my country include: clinical diagnosis, abnormal electrocardiogram, and determination of protein marker content. There are no typical changes, and the diagnostic coincidence rate of myocardial infarction is only 75% by relying solely on ECG changes and clinical symptoms. The detection of protein markers has the advantages of strong specificity and high sensitivity; therefore, the research on protein markers of myocardial infarction has been very active in recent years. Protein markers of myocardial infarction mainly include myoglobin, creatine kinase isoenzyme, and troponin I.
现有的蛋白标志物检测方法主要有酶联免疫法（ELISA）、免疫荧光法（IFA）和免疫胶体金法。ELISA和IFA具有特异性强、灵敏度高、重复性好的优点，但其操作程序复杂，只能在实验室进行；需要专业人员操作，用专门的仪器和设备，检测时间长，所需费用高。同时一次反应只能检测一个指标。免疫胶体金法测试简单快速、灵敏度高，成本低，且可以一次测试几种标志物。但其高灵敏度易导致假阳性结果，质量控制也不稳定。因此开发和使用联合检测以提高心肌梗塞诊断的灵敏度和特异度以及阳性预测值是现实迫切需要解决的课题。The existing protein marker detection methods mainly include enzyme-linked immunoassay (ELISA), immunofluorescence assay (IFA) and immunocolloidal gold method. ELISA and IFA have the advantages of strong specificity, high sensitivity, and good repeatability, but their operation procedures are complicated and can only be carried out in laboratories; professional personnel are required to operate, special instruments and equipment are used, the detection time is long, and the cost is high . Only one indicator can be detected in one reaction at the same time. The immunocolloidal gold method test is simple, fast, high sensitivity, low cost, and can test several markers at one time. However, its high sensitivity can easily lead to false positive results, and the quality control is not stable. Therefore, developing and using combined detection to improve the sensitivity, specificity and positive predictive value of the diagnosis of myocardial infarction is an urgent problem to be solved in reality.
发明内容Contents of the invention
本发明的目的在于克服现有技术存在的不足，而提供一种结构简单、制作方便、成本更低且灵敏度高的电流型心肌梗塞蛋白标记物的生物传感器薄层电极及其制备方法。The purpose of the present invention is to overcome the deficiencies in the prior art, and provide a thin-layer biosensor electrode for a current-type myocardial infarction protein marker with simple structure, convenient manufacture, lower cost and high sensitivity and a preparation method thereof.
本发明的目的是通过如下技术方案来完成的，一种快速检测心肌梗塞蛋白标记物的生物传感器薄层电极，它包括一块基板，基板上通过化学涂覆或电化学电镀工艺制备有至少一条形薄层电极，在所述薄层电极上制有至少一层含有与心肌梗塞蛋白标记物抗原相互作用、从而能够产生被检测电信号的抗体试剂层；所述抗体试剂层是通过无机凝胶层粘合在薄层电极表面。The object of the present invention is accomplished through the following technical scheme, a biosensor thin-layer electrode for rapid detection of myocardial infarction protein markers, which includes a substrate, on which at least one bar-shaped electrode is prepared by chemical coating or electrochemical plating process. A thin-layer electrode, on which at least one layer of an antibody reagent layer that interacts with the myocardial infarction protein marker antigen and can generate a detected electrical signal is formed; the antibody reagent layer is formed through an inorganic gel layer Bonded to the surface of the thin layer electrode.
作为优选：所述的薄层电极上面还粘合有纳米粒子含有层，并与抗体试剂层共同组成一叠层体；所述的纳米粒子含有层由电化学上为惰性的高分子化合物制成；所述的薄层电极为金属层、碳电极层以及银电极层之一。As a preference: the thin-layer electrode is also bonded with a nanoparticle-containing layer, and forms a laminate together with the antibody reagent layer; the nanoparticle-containing layer is made of an electrochemically inert polymer compound ; The thin layer electrode is one of metal layer, carbon electrode layer and silver electrode layer.
本发明利用心肌梗塞蛋白标记物抗原与生物传感器薄层电极上相应抗体相互作用，从而在相互作用过程中产生可以被检测的电信号；因此，本发明所述的生物传感器电电极，对于心肌梗塞蛋白标记物抗的检测是高特异性的，而且其检测的灵敏度非常高，重复性很好；具有结构简单、制作方便、成本更低且灵敏度高等特点。The present invention utilizes the myocardial infarction protein marker antigen to interact with the corresponding antibody on the biosensor thin-layer electrode, thereby generating an electrical signal that can be detected during the interaction process; therefore, the biosensor electric electrode described in the present invention is effective for myocardial infarction The detection of protein marker antibody is highly specific, and its detection sensitivity is very high, and the reproducibility is very good; it has the characteristics of simple structure, convenient production, lower cost and high sensitivity.
附图说明Description of drawings
图1是本发明所述薄层电极的正面结构示意图。Fig. 1 is a schematic view of the front structure of the thin-layer electrode of the present invention.
图2是本发明所述薄层电极的侧视结构示意图。Fig. 2 is a side view schematic diagram of the thin-layer electrode of the present invention.
具体实施方式Detailed ways
下面将结合附图对本发明作详细的介绍：图1、2所示，本发明所述的一种快速检测心肌梗塞蛋白标记物的生物传感器薄层电极，它包括一块基板1，基板1上通过化学涂覆或电化学电镀工艺制备有至少一条形薄层电极2，在所述薄层电极2上制有至少一层含有与心肌梗塞蛋白标记物抗原相互作用、从而能够产生被检测电信号的抗体试剂层3；所述抗体试剂层3是通过无机凝胶层4粘合在薄层电极2表面。The present invention will be described in detail below in conjunction with the accompanying drawings: shown in Fig. 1, 2, a kind of biosensor thin-layer electrode of the rapid detection myocardial infarction protein marker of the present invention, it comprises a substrate 1, passes through on the substrate 1 The chemical coating or electrochemical plating process is prepared with at least a strip-shaped thin-layer electrode 2, and at least one layer is formed on the thin-layer electrode 2 that contains a substance that interacts with the myocardial infarction protein marker antigen to generate a detected electrical signal. Antibody reagent layer 3 ; the antibody reagent layer 3 is bonded to the surface of the thin-layer electrode 2 through the inorganic gel layer 4 .
作为优选的一个实施例：所述的薄层电极2上面还粘合有纳米粒子含有层5，并与抗体试剂层3共同组成一叠层体；所述的纳米粒子含有层5由电化学上为惰性的高分子化合物制成；所述的薄层电极2为金属层、碳电极层以及银电极层之一。As a preferred embodiment: the thin-layer electrode 2 is also bonded with a nanoparticle-containing layer 5, and forms a laminate together with the antibody reagent layer 3; the nanoparticle-containing layer 5 is electrochemically formed. It is made of an inert polymer compound; the thin-layer electrode 2 is one of a metal layer, a carbon electrode layer and a silver electrode layer.
本发明通过化学镀覆、电化学电镀工艺制备的电极；可定性定量检测，重现性好，特异性高，使用寿命长，稳定性好的优点。The electrode prepared by the chemical plating and electrochemical plating process of the present invention has the advantages of qualitative and quantitative detection, good reproducibility, high specificity, long service life and good stability.
本发明属于对现有技术的改进，因此，上述未提及以及未公开的技术均为现有技术，且为本领域技术人员所熟练掌握，且在上述公开技术的基础上，能够方便的实施本发明。The present invention belongs to the improvement to prior art, therefore, above-mentioned unmentioned and undisclosed technology is prior art, and is mastered by those skilled in the art, and on the basis of above-mentioned disclosed technology, can implement conveniently this invention.
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