CN104297309A - Electrochemical transducer, and preparation method and application for detecting pathogenic bacteria - Google Patents
Electrochemical transducer, and preparation method and application for detecting pathogenic bacteria Download PDFInfo
- Publication number
- CN104297309A CN104297309A CN201410590803.7A CN201410590803A CN104297309A CN 104297309 A CN104297309 A CN 104297309A CN 201410590803 A CN201410590803 A CN 201410590803A CN 104297309 A CN104297309 A CN 104297309A
- Authority
- CN
- China
- Prior art keywords
- pathogenic bacteria
- oligonucleotide probe
- working fluid
- dispersion liquid
- detect
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention provides an electrochemical transducer, and a preparation method and application for detecting pathogenic bacteria, and relates to the field of pathogenic bacteria detection. The electrochemical transducer comprises a working electrode, wherein the working electrode is a wave carbon electrode with the surface modified with oligonucleotide probe 2. The electrochemical transducer further comprises a working solution 1 and a working solution 2, wherein the working solution 1 is a nanometer magnetic bead dispersion liquid marked with a pathogenic bacteria antibody; the working solution 2 is a gold nanometer particle dispersion liquid marked with a pathogenic bacteria antibody and oligonucleotide probe 1; the end 5 of the oligonucleotide probe 1 is modified with methylene blue, the end 3 of the oligonucleotide probe 2 is modified with amino, and the base sequences of the oligonucleotide probe 1 and the oligonucleotide probe 2 are complementary. According to the invention, the electrochemical transducer is convenient and rapid, and the detection sensitivity is significantly improved.
Description
Technical field
The present invention relates to pathogenic microbes detect field, be specifically related to a kind of detect pathogenic bacteria electrochemical sensor, preparation method and application thereof.
Background technology
Pathogenic bacteria (Pathogenic bacteria) refer to the microorganism that can cause disease.Pathogenic bacteria comprise bacterium, virus, conveyor screw, rickettsia, Chlamydia, mycoplasma, fungi and actinomyces etc.General said pathogenic bacteria refer to the bacterium in pathogenic microorganism.Pathogenic and its virulence of bacterium, invade quantity and portal of entry relevant.Although most bacterium is harmless even useful, a large portion can be caused a disease.Conditioned pathogen is only caused a disease under given conditions, bacterium can be allowed to enter blood if any wound, or when immunity reduces.
Such as: campylobacter jejuni is many animals as the bacterium that normally lives away from home of ox, sheep, dog and bird.A large amount of bacterium is had at their genital tract or enteron aisle, therefore by childbirth or manure contamination food and drinking-water.Campylobacter jejuni is a kind of infecting both domestic animals and human disease pathogen mattress, can cause humans and animals generation various diseases, and is a kind of borne Parasitic Encephalopathy cause of disease mattress, is considered to the main cause causing whole world human bacterial property to suffer from diarrhoea.Cardinal symptom is diarrhoea and stomachache, sometimes generates heat, and occasionally has vomiting and dehydration.The general susceptible of crowd, the incidence of disease of less than 5 years old children is the highest, and summer and autumn is common.Campylobacter jejuni has endotoxin to attack small intestine and colorectal mucosa causes chordapsus, also can cause outbreak of epidemic or the mass food poisoning of diarrhoea.Being generally 3 ~ 5 days latent period, is jejunum, ileum and colon to the pathogenic position of people.Colon bacillus (
e. coli) be commonly referred to Escherichia coli, be that Escherich found in 1885, be distributed in nature.Although most Escherichia coli and the mankind have good cooperation, still have the Escherichia coli of small part specific type to have great virulence, once infect, serious epidemic situation will be caused.Wherein most is representational is exactly the Escherichia coli that code name is O157:H7, and it is EHEC(enterohemorrhagic Escherichia coli) a member in family.After human infection EHEC, serious spasmodic colic and the hemorrhagic diarrhea of recurrent exerbation can be there is, simultaneously with performances such as heating, vomitings, caused by the toxin mostly being EHEC generation.Some severe infections person toxin causes hemolytic anemia with hematogenous spread, red blood cell, decrease of platelet; Acute renal failure also can be there is even dead when kidney is subject to involving.
The common methods of current detection pathogenic bacteria has: plating method, instrumental method, immuno-chromatographic test paper strip method etc.But these method complicated operations, to testing environment or instrument requirements high, consuming time longer, as time-consuming in flat band method and environmental requirement is strict, instrumental method not only costliness but also loaded down with trivial details.
Summary of the invention
The object of the present invention is to provide a kind of electrochemical sensor detecting pathogenic bacteria, can simply, fast, Sensitive Detection pathogenic bacteria.
Another object of the present invention is to provide the preparation method of the electrochemical sensor detecting pathogenic bacteria, and the method is simple, and efficiency is high.
Another object of the present invention is to provide the method adopting described electrochemical sensor to detect pathogenic bacteria, and the method is simple, quick, sensitive.
Object of the present invention adopts following technical scheme to realize.
Detect the electrochemical sensor of pathogenic bacteria, comprise working electrode, described working electrode is the ripple carbon electrode that finishing has oligonucleotide probe 2; Described sensor also comprises working fluid 1 and working fluid 2; Described working fluid 1 is for being marked with the nanometer magnetic bead dispersion liquid of anti-pathogenic bacteria antibody; Described working fluid 2 is for being marked with the golden nanometer particle dispersion liquid of anti-pathogenic bacteria antibody and oligonucleotide probe 1; 5 ' of described oligonucleotide probe 1 terminal modifiedly has methylenum careuleum, and 3 ' of oligonucleotide probe 2 terminal modifiedly has amino, the base sequence complementary of described oligonucleotide probe 1 and 2.
In the present invention, the nucleotide sequence (SEQ ID NO:1) of described oligonucleotide probe 1 is 5 '-GTGCACGGTACGCGAATCGG-3 ', nucleotide sequence (SEQ ID NO:2) the 5 '-CCGATTCGCGTACCGTGCAC-3 ' of described oligonucleotide probe 2.
In the present invention, described pathogenic bacteria can be campylobacter jejuni.
The present invention also provides the preparation method of the electrochemical sensor of described detection pathogenic bacteria, described finishing has the ripple carbon electrode of oligonucleotide probe 2 to adopt preparation with the following method: at ripple carbon electrodes deposited oxide graphene film, described graphene oxide film is modified oligonucleotide probe 2.
In the present invention, described finishing has the ripple carbon electrode of oligonucleotide probe 2 to adopt preparation with the following method: by the polishing of ripple carbon electrode alumina powder, in second alcohol and water, carry out ultrasonic cleaning; Graphene oxide suspension is dripped the ripple carbon electrodes after ultrasonic cleaning, obtains the ripple carbon electrode that surface deposition has graphene oxide film; Surface deposition has the ripple carbon electrode of graphene oxide film to activate in N-hydroxysuccinimide and 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride solution, then ripple carbon electrodes after activation drips oligonucleotide probe 2 solution, and 3 ' of described oligonucleotide probe 2 terminal modifiedly has amino, nucleotide sequence (SEQ ID NO:2) is 5 '-CCGATTCGCGTACCGTGCAC-3 '.
In the present invention, described working fluid 1 is adopted and is prepared with the following method: nanometer magnetic bead adopts 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and the activation of N-hydroxysuccinimide, then add anti-Antibodies of Campylobacter Jejuni to react, obtain the nanometer magnetic bead dispersion liquid being marked with anti-Antibodies of Campylobacter Jejuni.
In the present invention, described working fluid 2 is adopted and is prepared with the following method:
(1) in acetum, add three (2-carboxyethyl) phosphines and the 3 ' terminal modified bioprobe having sulfydryl, the 5 ' terminal modified methylenum careuleum, the nucleotide sequence (SEQ ID NO:1) of described bioprobe is 5 '-GTGCACGGTACGCGAATCGG-3 ', reacts;
(2) in step (1) reaction gained potpourri, add golden nanometer particle and react;
(3) in step (2) gained reaction mixture, add anti-Antibodies of Campylobacter Jejuni and react; Then deoxyadenosine triphosphate is added and bovine serum albumin(BSA) reacts;
(4) step (3) gained reaction mixture is centrifugal, gets precipitation and is resuspended in phosphate buffer, obtains the golden nanometer particle dispersion liquid being marked with anti-pathogenic bacteria antibody and oligonucleotide probe 1.
The present invention also provides above-mentioned electrochemical sensor to detect the method for pathogenic bacteria, comprises the steps:
(1) working fluid 1 is mixed with the solution containing variable concentrations pathogenic bacteria, be separated nanometer magnetic bead and be suspended in damping fluid, obtaining nanometer magnetic bead dispersion liquid;
(2) in nanometer magnetic bead dispersion liquid, add working fluid 2, react;
(3) be added drop-wise on working electrode by the potpourri that step (2) obtains and react, cleaning, apply voltage at working electrode two ends, sensed current signal, obtains the typical curve of pathogenic bacteria concentration and electric current;
(4) testing sample is detected according to step (1)-(3), the electric current obtained is substituted in typical curve, obtains the concentration of pathogenic bacteria in testing sample.
Compared with prior art, beneficial effect of the present invention is as follows:
Principle of the present invention as shown in Figure 1.Due to antigen and antibody mediated immunity identification, the nanometer magnetic bead that the present invention is marked with anti-pathogenic bacteria antibody and the golden nanometer particle being marked with anti-pathogenic bacteria antibody and oligonucleotide probe 1 all can pathogenic bacteria in specific identification sample, define magnetic bead-gold particle compound.Oligonucleotide probe 2 complementary pairing that the oligonucleotide probe 1 of magnetic bead-gold particle compound and working electrode surface are modified, the redox reaction of methylenum careuleum generation simultaneously, thus have the Transport And Transformation of electronics form electric current and be detected.The present invention adopts galvanochemistry as detection means, convenient and swift; Adopt the nanometer magnetic bead being marked with anti-pathogenic bacteria antibody as the means of enrichment pathogenic bacteria, significantly improve detection sensitivity.The present invention is universal method, only needs the monoclonal antibody of replacing specific recognition pathogenic bacteria, can realize quick, the Site Detection to any pathogenic bacteria, applied range, simply be easy to apply.
Accompanying drawing explanation
Fig. 1 shows the principle of detection method.
Fig. 2 is the typical curve of jejunum campylobacter bacteria concentration and strength of current.
Embodiment
In the present invention, room temperature refers to 20-25 DEG C.
Concentration is the PBS damping fluid (pH7.4) of 10 mM: get 137 mmolNaCl, 2.7 mmolKCl, 10 mmol Na
2hPO
4with 2 mmol KH
2pO4 is water-soluble, regulates pH to 7.4, is settled to 1L with water.
MES damping fluid: 100 mmol 2-(N-morpholines) ethyl sulfonic acid is water-soluble, and regulate pH to 5.9, be settled to 1 L with water.
If chemical reaction does not disclose temperature of reaction in the present invention, all refer at room temperature react.
Embodiment 1 detects the electrochemical sensor of campylobacter jejuni
(1) preparation of working fluid 1
Working fluid 1 is for being marked with the nanometer magnetic bead dispersion liquid of anti-campylobacter jejuni monoclonal antibody, and nanometer magnetic bead concentration is 10mmol/L, and the mol ratio of nanometer magnetic bead and anti-campylobacter jejuni monoclonal antibody is 1:6.
Get 1.08 g iron chloride and 0.2 g trisodium citrate is dissolved in 20 mL ethylene glycol, then add 1.2 g sodium acetates, stir 30 min, put in 200 DEG C of oil baths and be incubated 10 hours, obtain the nanometer magnetic bead dispersion liquid that concentration is 10 mM.
Get N-hydroxysuccinimide (NHS) the solution 15 μ L of 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC) solution 15 μ L and 0.01mM of nanometer magnetic bead solution 60 μ L, 0.1mM of 10 mM, mixing, react 10-30 minute under room temperature, nanometer magnetic bead is activated; Add the anti-campylobacter jejuni monoclonal antibody of 0.01 mg (purchased from abcam, production code member ab31468, anti-campylobacter jejuni antibody titer 1:200, anti-campylobacter jejuni monoclonal antibody affinity costant 1.2 × 10
10l/M), mix, under room temperature, react 2-4 hour, adopt magnetic frame magnetic resolution nanometer magnetic bead, be that the PBS damping fluid (pH7.4) of 10 mM is resuspended by concentration, obtain the nanometer magnetic bead dispersion liquid of anti-campylobacter jejuni labeling of monoclonal antibody.
(2) preparation of working fluid 2
Working fluid 2 is the golden nanometer particle dispersion liquids being marked with anti-campylobacter jejuni monoclonal antibody and oligonucleotide probe 1.In working fluid 2, golden nanometer particle concentration is 0.1 mg/mL, and anti-campylobacter jejuni monoclonal antibody, mol ratio between oligonucleotide probe 1 and golden nanometer particle are 1:1:10.5 ' of oligonucleotide probe 1 terminal modifiedly has methylenum careuleum, and nucleotides sequence is classified as 5 '-GTGCACGGTACGCGAATCGG-3 '.
The preparation method of golden nanometer particle dispersion liquid: the chlorauric acid solution adding 5 mL, 1 g/L in conical flask, then volume is supplied to 50 mL with aqua sterilisa, bumping is heated under stirring, adding 0.6 ~ 1 mL concentration is 1%(mass percentage concentration) sodium citrate solution, Keep agitation, stablize to claret to solution colour, obtaining concentration is 0.1 mg/mL golden nanometer particle dispersion liquid.
In vial, add aqueous acetic acid that 2 μ L concentration are 500 mM, bioprobe 1 that three (2-carboxyethyl) phosphine (TCEP) solution that 3 μ L concentration are 1mM and 20 μ L concentration are 1 nM, hybrid reaction 1 hour.Then the golden nanometer particle dispersion liquid (pH 8.0) that 200 μ L concentrate 10 times is added, hybrid reaction 1 hour.Then the anti-campylobacter jejuni monoclonal antibody (originating the same) that 100 μ L concentration are 0.1mg/mL is added, hybrid reaction 1 hour.Add deoxyadenosine triphosphate (dATP) solution and 20 μ L, 10%(w/v that 20 μ L concentration are 100 μMs) bovine serum albumin solution, hybrid reaction half an hour.By reaction mixture, under the centrifugal force of 9.4 g centrifugal 7 minutes, get precipitation and be resuspended in 500 μ LPBS damping fluids (10 mM, pH7.4), obtain the golden nanometer particle dispersion liquid being marked with anti-campylobacter jejuni monoclonal antibody and oligonucleotide probe 1, be positioned in 4 DEG C of environment, storage.
Bioprobe 1: at 3 ' the terminal modified sulfydryl of nucleotide fragments 5 '-GTGCACGGTACGCGAATCGG-3 ', the 5 ' terminal modified methylenum careuleum (MB), structural formula is MB-5 '-GTGCACGGTACGCGAATCGG-3 '-SH, is synthesized by Shanghai Sheng Gong biotech firm.
(3) preparation of working electrode
Working electrode is the ripple carbon electrode that finishing has oligonucleotide probe 2.
Ripple carbon electrode particle diameter is the alumina powder polishing of 0.05 micron, then in absolute ethyl alcohol and ultrapure water, distinguishes ultrasonic 2 minutes.
Taking 25 mg graphene oxides, to join 50 mL concentration be in the PBS solution of 0.1 mol/L, then make it be formed for ultrasonic 30 minutes even brown oxidation Graphene suspension that concentration is 0.5 mg/mL.
The above-mentioned graphene oxide suspension of 20 μ L is dripped in ripple carbon electrodes, dry, at ripple carbon electrodes deposited oxide graphene film.
There is by surface deposition the ripple carbon electrode of graphene oxide film to be placed in the NaOH solution that concentration is 1.5 mol/L, react 5 minutes under 1.5 V voltages.Take out ripple carbon electrode, the 2-(N-morpholine that 10 μ L contain 100 mmol/L N-hydroxysuccinimide (NHS) and 400 mmol/L 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC) is dripped at electrode surface) ethyl sulfonic acid (MES) damping fluid (0.1 mol/L, pH5.9), carry out priming reaction 1 hour, then use MES buffer solution for cleaning.
Electrode surface after activation drips the oligonucleotide probe 2 that 10 μ L concentration are 25 μMs, reacts 1 hour, with PBS damping fluid (10 mM, pH7.4) cleaning, obtains working electrode.
3 ' end of oligonucleotide probe 2 has decorations amino, and nucleotides sequence is classified as 5 '-CCGATTCGCGTACCGTGCAC-3 ', and structural formula is 5 '-CCGATTCGCGTACCGTGCAC-3 '-NH
2 +, synthesized by Shanghai Sheng Gong biotech firm.
(4) Ag/AgCl is contrast electrode, and platinum electrode is auxiliary electrode.
Embodiment 2 detects the campylobacter jejuni in actual sample
1. Criterion curve
The electrochemical sensor detecting campylobacter jejuni in embodiment 1 is adopted to detect campylobacter jejuni.Testing process all at room temperature.
(1) get 20 μ L working fluids 1 to join 1mL and be added with in the milk soln of variable concentrations campylobacter jejuni (ATCC33560, purchased from Central Plains, Beijing company), be placed on after mixing on rotary mixer, under 25 revs/min of conditions, react 10 minutes.Adopt magnetic frame magnetic resolution nanometer magnetic bead-campylobacter jejuni compound, remove supernatant, add PBS (pH 7.4) solution that 1mL concentration is 10 mM, and be placed on rotary mixer clean 3 minutes, repeated washing 3 times afterwards, finally add 100 μ L PBS damping fluids (10 mM, pH 7.4) resuspended, obtain nanometer magnetic bead-campylobacter jejuni compound dispersion liquid.
(2) respectively get 20 μ L immunomagnetic beads-campylobacter jejuni compound dispersion liquids and working fluid 2 mixes, react 30 minutes under 20 revs/min of conditions, obtain magnetic bead-gold particle complex solution.
(3) drip 10 μ L magnetic bead-gold particle complex solutions at working electrode surface, react 1 hour, clean with PBS damping fluid (10 mM, pH 7.4).
(4) Ag/AgCl is contrast electrode, and platinum electrode is auxiliary electrode, adopts Differential Pulse Voltammetry, applies 0.0 volt to-0.4 volt voltage, the electric current that testing electrode produces at electrode two ends.Obtain the typical curve (Fig. 2) of jejunum campylobacter bacteria concentration and strength of current, can find out that the sensing range of detection method is 5-400 cfu/mL.
2. concrete sample detection
Adopted by concrete sample the present embodiment title 1 method to detect, electric signal (electric current) value will be recorded and bring in mark song.
SEQUENCE LISTING
<110> Propagation and Food Test Center of Jiangsu Entry-Exit Inspection and Quarantine Bureau
<120> detects the electrochemical sensor of pathogenic bacteria, preparation method and application thereof
<130> 20141029
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 20
<212> DNA
<213> artificial
<220>
The nucleotide sequence of <223> oligonucleotide probe 1
<400> 1
gtgcacggta cgcgaatcgg 20
<210> 2
<211> 20
<212> DNA
<213> artificial
<220>
The nucleotide sequence of <223> oligonucleotide probe 2
<400> 2
ccgattcgcg taccgtgcac 20
Claims (8)
1. detect the electrochemical sensor of pathogenic bacteria, comprise working electrode, it is characterized in that described working electrode is the ripple carbon electrode that finishing has oligonucleotide probe 2; Described sensor also comprises working fluid 1 and working fluid 2; Described working fluid 1 is for being marked with the nanometer magnetic bead dispersion liquid of anti-pathogenic bacteria antibody; Described working fluid 2 is for being marked with the golden nanometer particle dispersion liquid of anti-pathogenic bacteria antibody and oligonucleotide probe 1; 5 ' of described oligonucleotide probe 1 terminal modifiedly has methylenum careuleum, and 3 ' of oligonucleotide probe 2 terminal modifiedly has amino, the base sequence complementary of described oligonucleotide probe 1 and 2.
2. detect the electrochemical sensor of pathogenic bacteria according to claim 1, it is characterized in that the nucleotides sequence of described oligonucleotide probe 1 is classified as 5 '-GTGCACGGTACGCGAATCGG-3 ', the nucleotide sequence 5 '-CCGATTCGCGTACCGTGCAC-3 ' of described oligonucleotide probe 2.
3. according to claim 1 or 2, detect the electrochemical sensor of pathogenic bacteria, it is characterized in that described pathogenic bacteria are campylobacter jejuni.
4. described in claim 1, detect the preparation method of the electrochemical sensor of pathogenic bacteria, it is characterized in that described finishing has the ripple carbon electrode of oligonucleotide probe 2 to adopt and prepares with the following method: at ripple carbon electrodes deposited oxide graphene film, described graphene oxide film is modified oligonucleotide probe 2.
5. detect the preparation method of the electrochemical sensor of pathogenic bacteria according to claim 4, it is characterized in that described finishing has the ripple carbon electrode of oligonucleotide probe 2 to adopt and prepares with the following method: by the polishing of ripple carbon electrode alumina powder, in second alcohol and water, carry out ultrasonic cleaning; Graphene oxide suspension is dripped the ripple carbon electrodes after ultrasonic cleaning, obtains the ripple carbon electrode that surface deposition has graphene oxide film; Surface deposition has the ripple carbon electrode of graphene oxide film to activate in N-hydroxysuccinimide and 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride solution, then ripple carbon electrodes after activation drips oligonucleotide probe 2 solution, and 3 ' of described oligonucleotide probe 2 terminal modifiedly has amino, nucleotides sequence is classified as 5 '-CCGATTCGCGTACCGTGCAC-3 '.
6. according to claim 4 or 5, detect the preparation method of the electrochemical sensor of pathogenic bacteria, it is characterized in that described working fluid 1 is adopted to prepare with the following method: nanometer magnetic bead adopts 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and the activation of N-hydroxysuccinimide, then add anti-Antibodies of Campylobacter Jejuni to react, obtain the nanometer magnetic bead dispersion liquid being marked with anti-Antibodies of Campylobacter Jejuni.
7. detect the preparation method of the electrochemical sensor of pathogenic bacteria according to claim 6, it is characterized in that described working fluid 2 is adopted and prepare with the following method:
(1) in acetum, add three (2-carboxyethyl) phosphines and the 3 ' terminal modified bioprobe having sulfydryl, the 5 ' terminal modified methylenum careuleum, the nucleotides sequence of described bioprobe is classified as 5 '-GTGCACGGTACGCGAATCGG-3 ', reacts;
(2) in step (1) reaction gained potpourri, add golden nanometer particle and react;
(3) in step (2) gained reaction mixture, add anti-Antibodies of Campylobacter Jejuni and react; Then deoxyadenosine triphosphate is added and bovine serum albumin(BSA) reacts;
(4) step (3) gained reaction mixture is centrifugal, gets precipitation and is resuspended in phosphate buffer, obtains the golden nanometer particle dispersion liquid being marked with anti-pathogenic bacteria antibody and oligonucleotide probe 1.
8. adopt the described electrochemical sensor of one of claim 1-3 to detect the method for pathogenic bacteria, comprise the steps:
(1) working fluid 1 is mixed with the solution containing variable concentrations pathogenic bacteria, be separated nanometer magnetic bead and be suspended in damping fluid, obtaining nanometer magnetic bead dispersion liquid;
(2) in nanometer magnetic bead dispersion liquid, add working fluid 2, react;
(3) be added drop-wise on working electrode by the potpourri that step (2) obtains and react, cleaning, apply voltage at working electrode two ends, sensed current signal, obtains the typical curve of pathogenic bacteria concentration and electric current;
(4) testing sample is detected according to step (1)-(3), the electric current obtained is substituted in typical curve, obtains the concentration of pathogenic bacteria in testing sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410590803.7A CN104297309B (en) | 2014-10-29 | 2014-10-29 | The detection electrochemical sensor of pathogenic bacteria, preparation method and applications |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410590803.7A CN104297309B (en) | 2014-10-29 | 2014-10-29 | The detection electrochemical sensor of pathogenic bacteria, preparation method and applications |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104297309A true CN104297309A (en) | 2015-01-21 |
| CN104297309B CN104297309B (en) | 2016-08-17 |
Family
ID=52317130
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410590803.7A Active CN104297309B (en) | 2014-10-29 | 2014-10-29 | The detection electrochemical sensor of pathogenic bacteria, preparation method and applications |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104297309B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109085222A (en) * | 2018-08-10 | 2018-12-25 | 青岛科技大学 | Ion liquid functionalization graphene vibrios DNA electrochemical sensor and its preparation method and application |
| CN109632910A (en) * | 2018-12-20 | 2019-04-16 | 温州大学 | Functionalization Fe3O4The preparation method of/Au nanocomposite and its application detected in mycotoxin |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060014172A1 (en) * | 2004-05-03 | 2006-01-19 | Nanosphere, Inc. | Aptamer-nanoparticle conjugates and method of use for target analyte detection |
| JP2007232675A (en) * | 2006-03-03 | 2007-09-13 | Matsushita Electric Ind Co Ltd | Gene detection method |
| CN101182580A (en) * | 2007-11-19 | 2008-05-21 | 中国科学院上海微***与信息技术研究所 | Gene or gene mutation measuring method based on magnetic beads and nanometer gold detecting probe |
| CN101250585A (en) * | 2008-03-28 | 2008-08-27 | 广州市搏克生物技术有限公司 | Method for detecting DNA, RNA and ultramicro-amount protein |
| CN101256191A (en) * | 2008-03-07 | 2008-09-03 | 中国科学院上海微***与信息技术研究所 | Method for determining minim proteins based on magnetic pearl and nano gold probe |
| CN101930002A (en) * | 2009-06-23 | 2010-12-29 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Hypersensitization technology for detecting pathogenic bacteria in water environment |
| CN101974624A (en) * | 2010-10-11 | 2011-02-16 | 福建国际旅行卫生保健中心 | Electrochemical DNA biosensor based novel legionnella detection method and electrochemical DNA biosensor |
-
2014
- 2014-10-29 CN CN201410590803.7A patent/CN104297309B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060014172A1 (en) * | 2004-05-03 | 2006-01-19 | Nanosphere, Inc. | Aptamer-nanoparticle conjugates and method of use for target analyte detection |
| JP2007232675A (en) * | 2006-03-03 | 2007-09-13 | Matsushita Electric Ind Co Ltd | Gene detection method |
| CN101182580A (en) * | 2007-11-19 | 2008-05-21 | 中国科学院上海微***与信息技术研究所 | Gene or gene mutation measuring method based on magnetic beads and nanometer gold detecting probe |
| CN101256191A (en) * | 2008-03-07 | 2008-09-03 | 中国科学院上海微***与信息技术研究所 | Method for determining minim proteins based on magnetic pearl and nano gold probe |
| CN101250585A (en) * | 2008-03-28 | 2008-08-27 | 广州市搏克生物技术有限公司 | Method for detecting DNA, RNA and ultramicro-amount protein |
| CN101930002A (en) * | 2009-06-23 | 2010-12-29 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Hypersensitization technology for detecting pathogenic bacteria in water environment |
| CN101974624A (en) * | 2010-10-11 | 2011-02-16 | 福建国际旅行卫生保健中心 | Electrochemical DNA biosensor based novel legionnella detection method and electrochemical DNA biosensor |
Non-Patent Citations (3)
| Title |
|---|
| M. BARREIROS DOS SANTOS等: "Detection of pathogenic Bacteria by Electrochemical Impedance Spectroscopy: Influence of the immobilization strategies on the sensor performance", 《PROCEDIA CHEMISTRY》 * |
| 王壮等: "空肠弯曲菌免疫磁珠富集方法的建立", 《药物生物技术》 * |
| 魏玲等: "4种食源性致病菌污染情况及其新型检测技术研究进展", 《食品科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109085222A (en) * | 2018-08-10 | 2018-12-25 | 青岛科技大学 | Ion liquid functionalization graphene vibrios DNA electrochemical sensor and its preparation method and application |
| CN109085222B (en) * | 2018-08-10 | 2020-07-28 | 青岛科技大学 | Ionic liquid functionalized graphene vibrio DNA electrochemical sensor and preparation method and application thereof |
| CN109632910A (en) * | 2018-12-20 | 2019-04-16 | 温州大学 | Functionalization Fe3O4The preparation method of/Au nanocomposite and its application detected in mycotoxin |
| CN109632910B (en) * | 2018-12-20 | 2021-09-07 | 温州大学 | Functionalized Fe3O4Preparation method of/Au nano composite material and application of Au nano composite material in mycotoxin detection |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104297309B (en) | 2016-08-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bhardwaj et al. | MOF–bacteriophage biosensor for highly sensitive and specific detection of Staphylococcus aureus | |
| Wang et al. | Efficient separation and quantitative detection of Listeria monocytogenes based on screen-printed interdigitated electrode, urease and magnetic nanoparticles | |
| Eissa et al. | Ultrasensitive peptide-based multiplexed electrochemical biosensor for the simultaneous detection of Listeria monocytogenes and Staphylococcus aureus | |
| CN105300963B (en) | For the preparation method and applications for the sandwich electrochemical luminescence immunosensor for detecting Marine Pathogenic Bacteria | |
| CN101504416A (en) | Novel methods for detecting bacillus coli by gold-coating magnetic granule in-situ initiating high-sensibility chemical luminescence | |
| WO2016005517A1 (en) | Methods for detecting target dna sequences | |
| CN108760855B (en) | Preparation method and application of graphene-polypyrrole-gold nanoparticle composite material | |
| Murasova et al. | Direct culture-free electrochemical detection of Salmonella cells in milk based on quantum dots-modified nanostructured dendrons | |
| CN105699471B (en) | A method of two kinds of substances are detected simultaneously | |
| CN110006971B (en) | Preparation method and application of aptamer sensor for detecting food-borne pathogenic bacteria through dual-channel output | |
| CN105891473B (en) | The preparation method and applications of food-borne pathogens immunosensor based on gold label silver stain signal amplification technique | |
| Jin et al. | NMR rapid detection of Salmonella in milk based on ultra-small iron oxide nanobiosensor | |
| CN104297309A (en) | Electrochemical transducer, and preparation method and application for detecting pathogenic bacteria | |
| Wang et al. | Perspectives for recognition and rapid detection of foodborne pathogenic bacteria based on electrochemical sensors | |
| Khosravi et al. | Preparation of immunomagnetic beads coupled with a rhodamine hydrazine immunosensor for the detection of Mycobacterium avium subspecies paratuberculosis in bovine feces, milk, and colostrum | |
| CN106053789B (en) | A kind of preparation method and application of the mycotoxin immunosensor based on graphite alkene toluidine blue composite | |
| Yu et al. | Saltatory rolling circle amplification-based ratiometric electrochemical biosensor for rapid detection of Salmonella enterica serovar typhimurium in food | |
| CN106872707A (en) | A kind of electrochemical immunosensor and its preparation and application for detecting zearalenone | |
| Mun et al. | Detection of Salmonella typhimurium by antibody/enzyme-conjugated magnetic nanoparticles | |
| CN104407132A (en) | Electrochemical sensor for detection of Escherichia coli and preparation method thereof | |
| CN105203756A (en) | Method for preparing quick magnetic separation electrochemistry immunosensor and method for detecting staphylococcus aureus | |
| CN101923092A (en) | Method for preparing carcinoembryonic antigen working electrode for screen printing electrode | |
| CN103207218B (en) | Electrochemical immunosensor making method and Streptococcus suis detection method using electrochemical immunosensor | |
| CN103983783B (en) | A kind of kit and application thereof detecting pathogenic bacteria | |
| Chen et al. | Antibiotic-based magnetic nanoprobes combined with mPCR for simultaneous detection of Staphylococcus aureus and Bacillus cereus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |