CN105861629A - Microbe concentration detection method based on immunomagnetic separation and biological catalysis - Google Patents
Microbe concentration detection method based on immunomagnetic separation and biological catalysis Download PDFInfo
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- CN105861629A CN105861629A CN201610183221.6A CN201610183221A CN105861629A CN 105861629 A CN105861629 A CN 105861629A CN 201610183221 A CN201610183221 A CN 201610183221A CN 105861629 A CN105861629 A CN 105861629A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
Abstract
The invention discloses a microbe concentration detection method based on immunomagnetic separation and biological catalysis. The method comprises the following steps: separating and enriching target microbes through a first bio-recognition element modified immunomagnetic element to improve the concentration and the purity of the target microbes; simultaneously modifying a nano-element with a second bio-recognition element and a predetermined enzyme, introducing an enzyme for catalyzing a biological reaction to a detection system in a sandwich manner, and changing the pH value of a catalysis solution by using the predetermined enzyme catalyzed biological reaction; and adding a pH indicator, and determining the content of the target microbes in the sample to be detected according to the color change of the pH indicator. The method integrates the purification and enrichment characteristics of immunomagnetic separation, the catalysis amplification characteristic of biological catalysis and the simple determination characteristic of the pH indicator, and also has the advantages of effective improvement of the detection sensitivity, shortening of the detection time, simplification of the detection process, and reduction of the detection cost.
Description
Technical field
The present invention relates to field of biological detection, be more particularly to a kind of based on immunity Magneto separate and the microorganism concn detection method of living things catalysis.
Background technology
Current China is faced with the food safety situation of sternness, and food safety affair takes place frequently.According to disease prevention and control center of country statistics, 2005-2014 China food poisoning is caused by microorganism more than 50%.Food safety prevention processed with discovery morning, early early warning and the morning that it is critical only that pathogenic microorganism that control, sets up pathogenic microorganism detection method sensitive, quick and has great importance.Although scientific research personnel has carried out substantial amounts of research in terms of improving microorganism detection sensitivity and reducing the detection time both at home and abroad, but realize the quickly detection of pathogenic microorganism in food and be still that a huge challenge, it is primarily due to pathogenic microorganism quantity in contaminated food the most less, complicated plus food substrate, conventional method of analysis is difficult to carry out pathogenic microorganism special detection highly sensitive, high.
Summary of the invention
(1) to solve the technical problem that
The technical problem to be solved in the present invention is how to improve the sensitivity of detection microorganism, reduction testing cost, shortening detection time.
(2) technical scheme
In order to solve above-mentioned technical problem, the invention provides a kind of based on immunity Magneto separate and the microorganism concn detection method of living things catalysis, said method comprising the steps of:
Described objective microbe is captured by S1, the immune magnetic element utilizing the first biological identification element of objective microbe to modify, form magnetics-objective microbe complex, and by magnetic field, described magnetics-objective microbe complex is separated, it is achieved the specific isolation enrichment of a small amount of objective microbe in sample；
S2, the nano-component utilizing the second biological identification element of predetermined enzyme and described objective microbe jointly to modify are combined with described magnetics-objective microbe complex, form magnetics-objective microbe-nano-component-multienzyme complex；
S3, utilize the described predetermined enzyme catalysis in described magnetics-objective microbe-nano-component-multienzyme complex corresponding with described predetermined enzyme treat catalytic solution, treat the pH value of catalytic solution described in change；
Treat the pH value of catalytic solution described in after S4, detection catalytic reaction, obtain the concentration of predetermined enzyme described in described magnetics-objective microbe-nano-component-multienzyme complex according to described pH value, determine the concentration of described objective microbe according to the concentration of described predetermined enzyme.
Preferably, described first biological identification element and the second biological identification element are different from the binding site of described objective microbe.
Preferably, described step S1 specifically includes following steps:
S11, utilize biotin, Streptavidin, magnetic material and described first biological identification element to prepare described immune magnetic element, described immune magnetic element is dissolved in phosphate buffer；
S12, carrying out mixing hatching with the sample solution containing described objective microbe by described immune magnetic element, described immune magnetic element captures described objective microbe, forms described magnetics-objective microbe complex；
S13, magnetic field is utilized to separate and be enriched with described magnetics-objective microbe complex from the mixed solution that described immune magnetic element is formed with described sample solution.
Preferably, described step S11 is prepared described immune magnetic element and specifically includes following steps:
S111, by described Streptavidin and magnetics coupling；
S112, by described biotin and described first biological identification element coupling；
First biological identification element of S113, the magnetics modified by described Streptavidin and described biotin modification carries out hatching and forms described immune magnetic element by the combination of biotin-Streptavidin.
Preferably, described nano-component is colloid JINHUA, and described predetermined enzyme is urase；
Described step S2 specifically includes following steps:
S21, utilization use seed mediated growth method to prepare colloid JINHUA；
S22, described colloid JINHUA is carried out described urase and the modification of described second biological identification element, form immune colloid gold flower, described immune colloid gold flower is dissolved in ultra-pure water；
S23, by described immune colloid gold flower carry out mixing hatching with the solution of described magnetics-objective microbe complex, form described magnetics-objective microbe-nano-component-multienzyme complex；
S24, magnetic field is utilized to isolate described magnetics-objective microbe-nano-component-multienzyme complex from the mixed solution that described immune colloid gold flower and the solution of described magnetics-objective microbe complex are formed.
Preferably, described step S4 specifically includes following steps:
S41, magnetic field is utilized to capture described magnetics-objective microbe-nano-component-multienzyme complex isolated supernatant.
S42, described supernatant is mixed with acid-base indicator, the color of described acid-base indicator changes, utilize the color intensity of variation of spectrogrph detection solution, obtain the concentration of urase in described magnetic material-object-colloid JINHUA-urase complex, thus obtain the concentration of described nano-component, the concentration of objective microbe is obtained according to the concentration of described nano-component.
Preferably, treat described in that catalytic solution is urea liquid；Described acid-base indicator is bromocresol purple.
After described step S24 further comprising the steps of:
S25, the sodium chloride solution (pH value is identical with bromocresol purple) that use contains tween successively and the washing of pure sodium chloride solution are to remove the described nano-component not being combined in the solution of described magnetics-objective microbe-nano-component-multienzyme complex with described objective microbe, the pH value of the wherein said sodium chloride solution containing tween is identical with the pH value of described bromocresol purple, to ensure that pH value is not affected by cleaning mixture.
Preferably, in described step S23, capture described magnetics-objective microbe complex first with magnetic field and remove supernatant, recycle described immune colloid gold flower and dissolve described magnetics-objective microbe complex, carry out mixing afterwards to hatch, form described magnetics-objective microbe-nano-component-multienzyme complex.
Preferably, described step S3 specifically includes following steps:
S31, utilize described urea liquid dissolve described magnetics-objective microbe-nano-component-multienzyme complex；
S32, the predetermined enzyme catalysis hydrolysis of urea in described magnetics-objective microbe-nano-component-multienzyme complex is utilized to react.
Preferably, in described step S31, ultra-pure water dissolved urea is utilized to form described urea liquid.
(3) beneficial effect
The invention provides a kind of based on immunity Magneto separate and the microorganism concn detection method of living things catalysis, the present invention first passes through the immune magnetic element of biological identification element modification and separates and be enriched with objective microbe, improve the concentration of objective microbe, predetermined enzymic catalytic reaction is utilized to change the pH value treating catalytic solution afterwards, effectively it is exaggerated detection signal, detection sensitivity can be increased substantially, and this enzymic catalytic reaction belongs to liquid--liquid reaction, reaction efficiency is higher than traditional solid-liquid phase reaction, detection speed can be increased substantially, the detection by quantitative result of available very small amount microorganism in 2 hours.Simultaneously, by enzyme catalysis, the present invention treats that the pH value of catalytic solution is treated in catalytic solution change, pH value must change can be observed intuitively by the change of acid-base indicator color or carry out quantitative observation with spectrogrph, without complicated equipment, the method hence with the present invention can be greatly lowered testing cost.
Accompanying drawing explanation
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, the accompanying drawing used required in embodiment or description of the prior art will be briefly described below, apparently, accompanying drawing in describing below is only some embodiments of the present invention, for those of ordinary skill in the art, on the premise of not paying creative work, it is also possible to obtain other accompanying drawing according to these accompanying drawings.
Fig. 1 be the present invention based on immunity Magneto separate and the flow chart of microorganism concn detection method of living things catalysis；
Fig. 2 be the present invention based on immunity Magneto separate and the operation principle schematic diagram of microorganism concn detection method of living things catalysis；
Fig. 3 is the full spectrogram of the solution after variable concentrations sal volatile and bromocresol purple are hatched in the present invention；
Fig. 4 is the light intensity value under characteristic wavelength of the solution after variable concentrations sal volatile and bromocresol purple are hatched in the present invention.
Detailed description of the invention
With embodiment, the present invention is described in further detail below in conjunction with the accompanying drawings.Following example are used for illustrating the present invention, but can not be used for limiting the scope of the present invention.
A kind of based on immunity Magneto separate and the microorganism concn detection method of living things catalysis, as it is shown in figure 1, said method comprising the steps of:
Described objective microbe is captured by S1, the immune magnetic element utilizing the first biological identification element of objective microbe to modify, form magnetics-objective microbe complex, and by magnetic field, described magnetics-objective microbe complex carried out sharp separation, thus realize the specific isolation enrichment of a small amount of pathogenic bacterium in sample；
S2, the nano-component utilizing the second biological identification element of predetermined enzyme and described objective microbe jointly to modify are combined with described magnetics-objective microbe complex, form magnetics-objective microbe-nano-component-multienzyme complex；
S3, utilize the described predetermined enzyme catalysis in described magnetics-objective microbe-nano-component-multienzyme complex corresponding with described predetermined enzyme treat catalytic solution, treat the pH value of catalytic solution described in change；
Treat the pH value of catalytic solution described in after S4, detection catalytic reaction, obtain the concentration of predetermined enzyme described in described magnetics-objective microbe-nano-component-multienzyme complex according to described pH value, determine the concentration of described objective microbe according to the concentration of described predetermined enzyme.
In said method, immunity magnetic separation technique has simple to operate, separation efficiency advantages of higher, its cardinal principle is the magnetic material utilizing and being modified with biological identification element (antibody etc.), specifically identify the target pathogenic bacterium in food substrate, and by magnetic field, magnetic material carried out sharp separation, thus realize the specific isolation enrichment of a small amount of pathogenic bacterium in food samples.Meanwhile, said method has the immunity purification enrichment feature of Magneto separate, the catalysis amplification feature of living things catalysis and the simple mensuration feature of acid-base indicator concurrently, can be effectively improved detection sensitivity, reduce the detection time, simplify detection process, reduce testing cost.
Further, described step S1 specifically includes following steps:
S11, utilize biotin, Streptavidin, magnetic material and described first biological identification element to prepare described immune magnetic element, described immune magnetic element is dissolved in phosphate buffer；
S12, carrying out mixing hatching with the sample solution containing described objective microbe by described immune magnetic element, described immune magnetic element captures described objective microbe, forms described magnetics-objective microbe complex；
S13, magnetic field is utilized to separate and be enriched with described magnetics-objective microbe complex from the mixed solution that described immune magnetic element is formed with described sample solution.
Described step S11 is prepared described immune magnetic element and specifically includes following steps:
S111, by described Streptavidin and magnetics coupling；
S112, by described biotin and described first biological identification element coupling；
First biological identification element of S113, the magnetics modified by described Streptavidin and described biotin modification carries out hatching and forms described immune magnetic element by the combination of biotin-Streptavidin.
Further, described first biological identification element and the second biological identification element are different from the binding site of described objective microbe, to ensure that the first biological identification element and the second biological identification element can be combined with objective microbe by different binding sites simultaneously, the combination rate of the other side that is independent of each other.
Preferably, described first biological identification element is monoclonal antibody, described magnetics is magnetic bead, described immune magnetic element is immunomagnetic beads, will be clear that the present invention does not limit the first biological identification element as monoclonal antibody, can also select other bio-identification material according to practical situation, similarly the present invention does not limit magnetics as magnetic bead, it is also possible to select other magnetic material according to practical situation.
Described nano-component is but not limited to colloid JINHUA, and described predetermined enzyme is but not limited to urase, described in treat that catalytic solution is but not limited to urea liquid；Described acid-base indicator is but not limited to bromocresol purple, specifically can choose according to practical situation.
Use the biological identification element specific isolation objects such as antibody, have and preferably detect specificity, it is possible to reduce the interference effect of other molecule in sample background；Use specificity preferable urase catalyzing urea to produce ammonium radical ion and carbanion, the interference effect of other molecule can be reduced.
Further, described step S2 specifically includes following steps:
S21, utilization use seed mediated growth method to prepare colloid JINHUA；
S22, described colloid JINHUA is carried out described urase and the modification of described second biological identification element, form described immune colloid gold flower, described immune colloid gold flower is dissolved in ultra-pure water；
S23, by described immune colloid gold flower carry out mixing hatching with the solution of described magnetics-objective microbe complex, form described magnetics-objective microbe-nano-component-multienzyme complex；
S24, magnetic field is utilized to isolate described magnetics-objective microbe-nano-component-multienzyme complex from the mixed solution that described immune colloid gold flower and the solution of described magnetics-objective microbe complex are formed.
S25, the sodium chloride solution (pH value is identical with bromocresol purple) that use contains tween successively and the washing of pure sodium chloride solution are to remove the described nano-component not being combined in the solution of described magnetics-objective microbe-nano-component-multienzyme complex with described objective microbe, the pH value of the wherein said sodium chloride solution containing tween is identical with the pH value of described bromocresol purple, to ensure that pH value is not affected by cleaning mixture.
Preferably, in described step S23, capture described magnetics-objective microbe complex first with magnetic field and remove supernatant, recycle described immune colloid gold flower and dissolve described magnetics-objective microbe complex, carry out mixing afterwards to hatch, form described magnetics-objective microbe-nano-component-multienzyme complex.Magnetic field rich magnetic element-objective microbe complex is utilized to further increasing the degree of accuracy of detection.
In step S2, washed by Magneto separate and different solutions and remove unconjugated colloid JINHUA and do not change the pH of complex, reduce background noise, improve system signal noise ratio, improve accuracy of detection.
Further, described step S3 specifically includes following steps:
S31, utilize described urea liquid dissolve described magnetics-objective microbe-nano-component-multienzyme complex；
S32, the predetermined enzyme catalysis hydrolysis of urea in described magnetics-objective microbe-nano-component-multienzyme complex is utilized to react.
Wherein, step S31 utilize ultra-pure water dissolved urea form described urea liquid.
Further, described step S4 specifically includes following steps:
S41, magnetic field is utilized to capture described magnetics-objective microbe-nano-component-multienzyme complex isolated supernatant.
S42, described supernatant is mixed with acid-base indicator, the color of described acid-base indicator changes, utilize the color intensity of variation of spectrogrph detection solution, obtain the concentration of urase in described magnetic material-object-colloid JINHUA-urase complex, thus obtain the concentration of described nano-component, the concentration of objective microbe is obtained according to the concentration of described nano-component.It should be noted that, in step S42, it is also possible to the concentration of objective microbe is directly determined by the concentration of urase.
Owing to enzyme catalysis has amplification signal, it is thus possible to improve the effect of detection sensitivity, be widely used detection field, the present invention utilizes urase catalyzing urea to produce carbonate and ammonium radical ion, urea liquid pH value is caused to raise and change the principle of the color of acid-base indicator-bromocresol purple, it is achieved the detection to microorganism.Fig. 3 is the full spectrogram of the solution after variable concentrations sal volatile and bromocresol purple are hatched in the present invention, Fig. 4 is the light intensity value under characteristic wavelength of the solution after variable concentrations sal volatile and bromocresol purple are hatched in the present invention, matched curve is: y=4384.4ln (x)+123.6, wherein: x is ammonium carbonate concentration, y is light intensity, R2It is 0.98.According to Fig. 3 and Fig. 4, utilize the color change of bromocresol purple to obtain the concentration of ammonium carbonate, so that it is determined that the concentration of urase, i.e. can determine that the concentration of objective microbe according to the concentration of urase.
Fig. 2 be the present invention based on immunity Magneto separate and the operation principle schematic diagram of microorganism concn detection method of living things catalysis, as shown in Figure 2, the present invention first passes through monoclonal antibody or the immunomagnetic ca pture objective microbe of other bio-identification material modification, form magnetics-objective microbe complex, such as Listeria monocytogenes, utilize magnetic field separation afterwards and be enriched with objective microbe, improve concentration and the purity of objective microbe；Then, colloid JINHUA or other nano material are modified polyclonal antibody or another kind of bio-identification material and urase or the enzyme of other identity function simultaneously, be combined with magnetics-objective microbe complex, in detection system, introduce urase by double antibodies sandwich mode, utilize urase catalyzing urea hydrolysis to change the pH value of urea liquid；Finally, add bromocresol purple or other acid-base indicator, and utilize objective microbe content in the color change detection sample to be tested of acid-base indicator.The method has the immunity purification enrichment feature of Magneto separate, the catalysis amplification feature of urase and the simple mensuration feature of acid-base indicator concurrently, can be effectively improved detection sensitivity, reduce the detection time, simplify detection process, reduce testing cost.
Embodiment of above is merely to illustrate the present invention, rather than limitation of the present invention.Although the present invention being described in detail with reference to embodiment, it will be understood by those within the art that, technical scheme is carried out various combination, amendment or equivalent, without departure from the spirit and scope of technical solution of the present invention, all should contain in the middle of scope of the presently claimed invention.
Claims (10)
1. based on immunity Magneto separate and a microorganism concn detection method for living things catalysis, its
It is characterised by, said method comprising the steps of:
S1, utilize objective microbe first biological identification element modify immune magnetic element pair
Described objective microbe captures, and forms magnetics-objective microbe complex, and passes through
Described magnetics-objective microbe complex is separated by magnetic field, it is achieved a small amount of mesh in sample
The specific isolation enrichment of mark microorganism；
S2, the second biological identification element of predetermined enzyme and described objective microbe is utilized jointly to modify
Nano-component be combined with described magnetics-objective microbe complex, formed magnetic unit
Part-objective microbe-nano-component-multienzyme complex；
S3, utilize in described magnetics-objective microbe-nano-component-multienzyme complex described
What predetermined enzyme catalysis was corresponding with described predetermined enzyme treats catalytic solution, treats catalytic solution described in change
PH value；
Treat the pH value of catalytic solution described in after S4, detection catalytic reaction, obtain according to described pH value
To the concentration of predetermined enzyme described in described magnetics-objective microbe-nano-component-multienzyme complex,
Concentration according to described predetermined enzyme determines the concentration of described objective microbe.
Method the most according to claim 1, it is characterised in that described first bio-identification
Element and the second biological identification element are different from the binding site of described objective microbe.
Method the most according to claim 1, it is characterised in that described step S1 is concrete
Comprise the following steps:
S11, utilize biotin, Streptavidin, magnetic material and described first bio-identification
Described immune magnetic element prepared by element, and described immune magnetic element is dissolved in phosphate-buffered
Liquid；
S12, described immune magnetic element is carried out with the sample solution containing described objective microbe
Mixing is hatched, and described immune magnetic element captures described objective microbe, forms described magnetic unit
Part-objective microbe complex；
S13, utilize magnetic field molten with the mixing that described sample solution is formed from described immune magnetic element
Liquid separates and is enriched with described magnetics-objective microbe complex.
Method the most according to claim 3, it is characterised in that make in described step S11
Standby described immune magnetic element specifically includes following steps:
S111, by described Streptavidin and magnetics coupling；
S112, by described biotin and described first biological identification element coupling；
The of S113, the magnetics that described Streptavidin is modified and described biotin modification
One biological identification element carries out hatching and forms described immunity by the combination of biotin-Streptavidin
Magnetics.
5. according to the method described in any one of Claims 1-4, it is characterised in that described in receive
Rice element is colloid JINHUA, and described predetermined enzyme is urase；
Described step S2 specifically includes following steps:
S21, utilization use seed mediated growth method to prepare colloid JINHUA；
S22, described colloid JINHUA is carried out described urase and described second biological identification element is repaiied
Decorations, form immune colloid gold flower, and described immune colloid gold flower is dissolved in ultra-pure water；
S23, by described immune colloid gold flower with described magnetics-objective microbe complex
Solution carries out mixing and hatches, and forms described magnetics-objective microbe-nano-component-enzyme and is combined
Thing；
S24, utilize magnetic field from described immune colloid gold flower with described magnetics-objective microbe
The mixed solution that the solution of complex is formed is isolated described magnetics-objective microbe-receive
Rice element-multienzyme complex.
Method the most according to claim 5, it is characterised in that described step S4 is concrete
Comprise the following steps:
S41, utilize magnetic field capture described magnetics-objective microbe-nano-component-multienzyme complex
And isolated supernatant；
S42, being mixed with acid-base indicator by described supernatant, the color of described acid-base indicator is sent out
Changing, utilizes the color intensity of variation of spectrogrph detection solution, obtains described magnetic material-mesh
The concentration of the urase in mark thing-colloid JINHUA-urase complex, thus obtain described nano-component
Concentration, obtains the concentration of objective microbe according to the concentration of described nano-component.
Method the most according to claim 6, it is characterised in that described in treat that catalytic solution is
Urea liquid；Described acid-base indicator is bromocresol purple；
After described step S24 further comprising the steps of:
S25, the successively use sodium chloride solution (pH value is identical with bromocresol purple) containing tween
Wash with pure sodium chloride solution to remove described magnetics-objective microbe-nano-component-enzyme
The described nano-component not being combined with described objective microbe in the solution of complex, wherein said
The pH value of the sodium chloride solution containing tween is identical with the pH value of described bromocresol purple, to ensure
PH value is not affected by cleaning mixture.
Method the most according to claim 7, it is characterised in that in described step S23,
Capture described magnetics-objective microbe complex first with magnetic field and remove supernatant, then
Utilize described immune colloid gold flower dissolve described magnetics-objective microbe complex, laggard
Row mixing is hatched, and forms described magnetics-objective microbe-nano-component-multienzyme complex.
Method the most according to claim 8, it is characterised in that described step S3 is concrete
Comprise the following steps:
S31, utilize described urea liquid dissolve described magnetics-objective microbe-nano-component-
Multienzyme complex；
S32, utilize in described magnetics-objective microbe-nano-component-multienzyme complex predetermined
Enzyme catalysis hydrolysis of urea reacts.
Method the most according to claim 9, it is characterised in that in described step S31,
Ultra-pure water dissolved urea is utilized to form described urea liquid.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107202880A (en) * | 2017-05-24 | 2017-09-26 | 中国农业大学 | A kind of microorganism detection method analyzed based on electrochemical impedance phase angle |
| CN110501494A (en) * | 2019-07-16 | 2019-11-26 | 中国农业大学 | A kind of microorganism detection method based on manganese dioxide nano flower and fluorescent material |
| CN113933364A (en) * | 2021-09-03 | 2022-01-14 | 中国科学院上海微系统与信息技术研究所 | Target concentration detection method based on silicon nanowire field effect biochemical sensor |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104833802A (en) * | 2015-04-24 | 2015-08-12 | 中国农业大学 | Microbial detection method based on immunomagnetic separation and urease catalysis |
-
2016
- 2016-03-28 CN CN201610183221.6A patent/CN105861629A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104833802A (en) * | 2015-04-24 | 2015-08-12 | 中国农业大学 | Microbial detection method based on immunomagnetic separation and urease catalysis |
Non-Patent Citations (1)
| Title |
|---|
| CHEN Q等: "A sensitive impedance biosensor based on immunomagnetic separation and urease catalysis for rapid detection of Listeria monocytogenes using an immobilization-free interdigitated array microelectrode", 《BIOSENSORS AND BIOELECTRONICS》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107202880A (en) * | 2017-05-24 | 2017-09-26 | 中国农业大学 | A kind of microorganism detection method analyzed based on electrochemical impedance phase angle |
| CN110501494A (en) * | 2019-07-16 | 2019-11-26 | 中国农业大学 | A kind of microorganism detection method based on manganese dioxide nano flower and fluorescent material |
| CN110501494B (en) * | 2019-07-16 | 2020-10-09 | 中国农业大学 | Microorganism detection method based on manganese dioxide nanoflowers and fluorescent materials |
| CN113933364A (en) * | 2021-09-03 | 2022-01-14 | 中国科学院上海微系统与信息技术研究所 | Target concentration detection method based on silicon nanowire field effect biochemical sensor |
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