CN107037204A - A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes - Google Patents

A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes Download PDF

Info

Publication number
CN107037204A
CN107037204A CN201710200617.1A CN201710200617A CN107037204A CN 107037204 A CN107037204 A CN 107037204A CN 201710200617 A CN201710200617 A CN 201710200617A CN 107037204 A CN107037204 A CN 107037204A
Authority
CN
China
Prior art keywords
nano
coalo
probes
bacteria
anaerobic bacteria
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.)
Pending
Application number
CN201710200617.1A
Other languages
Chinese (zh)
Inventor
张锦胜
万益琴
彭红
巫小丹
邹灯超
郑洪立
赵茹
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanchang University
Original Assignee
Nanchang University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Nanchang University filed Critical Nanchang University
Priority to CN201710200617.1A priority Critical patent/CN107037204A/en
Publication of CN107037204A publication Critical patent/CN107037204A/en
Pending legal-status Critical Current

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N24/00Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
    • G01N24/08Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
    • G01N24/088Assessment or manipulation of a chemical or biochemical reaction, e.g. verification whether a chemical reaction occurred or whether a ligand binds to a receptor in drug screening or assessing reaction kinetics
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56911Bacteria

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Physics & Mathematics (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Food Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • High Energy & Nuclear Physics (AREA)
  • Toxicology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Virology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes, belongs to tcm development applied technical field.The present invention dependent on set up based on prepares CoAlO nano-probes be used for screen suppression anaerobic bacteria Chinese medicine preparation method.The characteristics of paramagnetic CoAlO nano-probes prepared can be specifically bound to object bacteria is coated with using antibody, the influence using CoAlO paramagnetic properties to the NMR relaxation time detects whether contain object bacteria in sample.Paramagnetic CoAlO nano-probes, NMR relaxation time showing outlet sexual intercourse within the specific limits, i.e. nano Co AlO contents are big, the nuclear magnetic resonance spin Lattice Relaxation Time and transverse relaxation time value of sample are smaller, detection object bacteria can be quantified in certain limit, so that the fungistatic effect of indirect assessment Chinese medicine preparation.This method can be used for the recipe determination for having the Chinese medicine of inhibitory action to anaerobic bacteria, accelerate the development rate of tcm product.

Description

A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes
Technical field
The present invention relates to a kind of screening technique of Chinese medicine preparation.
Background technology
Antibiotic simultaneously, will also result in human body infringement, influence liver, renal function, gastrointestinal reaction etc. in sterilization.Abuse is anti- Raw element may cause the generation of some bacterial resistance phenomenons, and the treatment to infection can become very difficult.At present, almost without one Plant antibacterials and resistance phenomenon is not present.Chinese medicine and formula of the exploitation with antibacterial action have positive effect.
Many Chinese medicines and magistral medicines have antibacterial and anti-inflammation functions, and some medium-height grass the effective elements of the medicines and molecular structure etc. are also Through studying clear whole or in part.The main component berberine of such as coptis and golden cypress stop dysentery(Berberine), radix scutellariae antibacterial Main component baicalein etc..A host of facts prove, the medicine that ancient Chinese working people is accumulated by long-term practice Legacy is extremely abundant, extremely valuable.We should value the great treasure-house of this motherland's medicine and pharmacology, make great efforts to excavate, be subject to Improve, can therefrom develop the Chinese medicine with antibacterial and anti-inflammation functions, can replace or part substitution antibiotic.
Anaerobic bacteria be a class under anaerobic than the bacterium grown in aerobic environment, and can not be with the presence of air In the case of the bacterium that grows.This kind of bacterium lacks complete metabolism enzyme system, and its energetic supersession is carried out in the way of anaerobic fermentation. It can cause the infection of human body different parts, including appendicitis, cholecystitis, tympanitis, mouth infection, endocarditis, osteomyelitis, A variety of diseases such as peritonitis.It is usually in sterilizing in Chinese medicine new formulation procedures of traditional exploitation for certain anaerobic bacteria Traditional Chinese medicinal components are added after inoculation of medium anaerobic bacteria, are cultivated in special anaerobic culture box.Sampled again after a period of time into Row line culture, calculates clump count, so that it is determined that whether certain Chinese medicine compositing formula has bacteriostasis.Line culture is also required to detesting Cultivated in oxygen incubator, and cultivation cycle at least takes several days, therefore whole cycle is long, efficiency is low.This method is asked for above-mentioned Inscribe a kind of rapid screening method of Chinese medicine preparation of invention.
The general principle of this method:Monoclonal antibody or antigen molecule are by Covalent bonding together, and this combination will not change Monoclonal antibody, the immunological characteristic of antigen and bioactivity, specific monoclonal antibody only can be with specific antigen knots Close.And quaternary CoAlO materials have ferromagnetism, its Elements C o outer layers have multiple unpaired electrons, strong with high saturation Degree, particle diameter it is small to a certain extent when, it may appear that paramagnetic and superparamagnetic characteristic CoAlO nano materials can be to the cores of surrounding water molecules The influence of magnetic resonance spins-Lattice Relaxation Time is very big, the effect with amplification nuclear magnetic resonance relaxation signals.It is very micro The NMR relaxation time that CoAlO nano materials will result in water declines to a great extent.Therefore can be by building CoAlO nanometers of spies Pin, detects whether there is object bacteria from the angle of magnetic resonance, so that the bacteriostasis of indirect assessment traditional Chinese medicine ingredients.
Key step:
1)Prepared by CoAlO nano-probes.Commercially paramagnetic CoAlO nano materials, can also be by other method system Standby nano level CoAlO.Using silane coupler, its formula is:Y(CH2)nSiX3.Herein, n is 0-3;X is hydrolyzable base Group;Y is organo-functional group.X is typically chloro, methoxyl group, ethyoxyl, acetoxyl group etc., and silicon is generated when these groups are hydrolyzed Alcohol (Si (OH)3), and combined with inorganic substances, form siloxanes.Y be vinyl, amino, epoxy radicals, methacryloxy, Sulfydryl.These reactive groups can react and combine with organic substance.Therefore, can be in inorganic substances by using silane coupler " molecular bridge " is erected between the interface of organic substance, the material of two kinds of property great disparities is linked together and improves composite The effect of performance and increase adhesive strength.The amination or carboxylated of surface modification can be obtained by being reacted by silane coupler CoAlO nano materials.Further by coupled antibody, surface-functionalized, formation specific immunity probe can be achieved, it is unnecessary to close Avtive spot, excessive antibody can be separated after centrifugation, you can to obtain the CoAlO nano-probes of resistance modification.
2)The sample cell made using polystyrene material carries out Chinese medicine screening.Polystyrene material and ELISA Plate class Seemingly, detecting probe surface antibody can be combined on the surface of polystyrene material due to suction-operated, still, and the antibody of detecting probe surface is inhaled The speed of polystyrene material surface is attached to compared with the speed of detecting probe surface antibody binding object bacteria, the former is slow-footed more, Both are competitive relations.Therefore, the culture medium of the anaerobic bacteria of sterilizing, vaccination target anaerobic bacteria are added in polystyrene sample pipe The Chinese medicine extract for needing to screen is added afterwards, after closing anaerobic culture for a period of time, adds step 1)Prepared CoAlO nanometers Probe, is sufficiently mixed concussion reaction for a period of time, if there is target bacteria growing in sample cell, CoAlO nano-probes surface it is anti- Body can be combined with object bacteria first, so that CoAlO nano-probes are stayed in a liquid.If there is no object bacteria in sample cell, The antibody on CoAlO nano-probes surface can be adsorbed gradually in polystyrene sample pipe surface, so that not having CoAlO to receive in liquid Rice probe.Preceding to have addressed, the NMR relaxation time that micro CoAlO nano materials will result in water declines to a great extent.With nothing The medium liquid without vaccination target bacterium of bacterium is blank, and the relaxation time of the suspension measured is compared and significantly reduced, then Illustrate to contain probe in liquid, so as to there is object bacteria in indirect proof sample, the amount of probe and the drop-out value in relaxation time are in just Than the amount of object bacteria can be gone out with indirect quantification by the quantitative probe of mark-on.With the presence of object bacteria, then illustrate Traditional Chinese medicine bacteriostatic effect not Substantially, it is on the contrary then explanation have bacteriostasis.
The major advantage of this method is exactly quick, sensitivity height.Object bacteria can be realized and continuously existed to a certain extent Line is monitored.Time of even several days 2-3 days relative to anaerobic bacteria culture, magnetic resonance detection only needs a few minutes.The method is not required to Anaerobic culture box is wanted, can cultivate the sample seal of tube after need to being only inoculated with anaerobic, will be quantitative with syringe during detection Probe injection can carry out follow-up magnetic resonance detection.Therefore, the screening of extensive Chinese medicine and formula can be made of this method.
The content of the invention
A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes.This method is a kind of objective effective Detection sample in object bacteria method, the quick screening of extensive Chinese medicine and formula can be done, so that big to a certain extent Big reduction is directed to certain anaerobic bacteria effective tcm development time.
A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes.Using NMR to paramagnetic The response sensibility of material, proposes NMR relaxation Parameters variation and the phase of CoAlO nano-particle paramagnetic immunological probe contents Closing property index.This method is matched somebody with somebody dependent on the detection that paramagnetic nano CoAlO nano-probes are used for object bacteria in sample to assess Chinese medicine The fungistatic effect of side., can be to special in sample using the paramagnetic CoAlO nano-probes for being coupled monoclonal antibody specific Property anaerobic bacteria is detected.Because NMR is very sensitive to CoAlO nano-particles, there is micro paramagnetic CoAlO and receive Rice corpuscles, then the spin-lattice relaxation time of water and/spin-spin relaxation will be remarkably decreased.Pass through blank control and mark-on The immunological probe content in sample is quantitatively detected, the anaerobic bacteria clump count gone out so as to indirect quantification in sample.Detect Anaerobism bacterial content and probe content linear correlation, degree of fitting is preferable.Finally between paramagnetic CoAlO nano-probes and anaerobic bacteria Corresponding relation is tie, determines the anaerobic bacteria clump count in sample.So as to determine whether the bacteriostasis of Chinese medicine is effective indirectly.Phase For the anaerobic bacteria culture time of even several days 2-3 days, magnetic resonance detection only needs a few minutes.Therefore, it can be made greatly of this method The screening of scale Chinese medicine and formula.
The present invention is achieved in that step is as follows:
1)Target bacteria antibody is coated with the preparation of paramagnetic nano CoAlO nano-probes.
2)The culture medium of the anaerobic bacteria of sterilizing, vaccination target anaerobic bacteria are added in polystyrene sample pipe.Adding needs The Chinese medicine extract of screening, after culture a period of time, after culture a period of time, adds step 1)Prepared CoAlO nanometers are visited Pin, is sufficiently mixed concussion reaction for a period of time, if there is object bacteria in sample cell, first can be with CoAlO nano-probes surface Antibody binding, so that CoAlO nano-probes are stayed in a liquid.If not there is object bacteria in sample cell, CoAlO nanometers of spies The antibody of wire surface can gradually be adsorbed and separated in polystyrene sample pipe surface with liquid.Now, nuclear-magnetism is carried out to sample cell The measure in resonance relaxation time.Using the sterile medium liquid without vaccination target bacterium as blank, the relaxation of the suspension measured The Henan time, then explanation contained probe compared to significantly reducing, so that have object bacteria in indirect proof sample, the amount and relaxation of probe The decline of time is proportional, and by mark-on quantitative probe, indirect quantification goes out the amount of object bacteria.
The CoAlO nano-probes are the nanoscale CoAlO materials with paramagnetic properties, and nanometer particle size is less than 300 nanometers.
The change in final relaxation time of the detection evaluation method based on nuclear magnetic resonance technique of described object bacteria, during relaxation Between include spin-lattice relaxation time and spin spin relaxation time.
Beneficial effects of the present invention:The present invention, which provides a kind of objectively quick screening, the Chinese medicine of inhibitory action to anaerobic bacteria The method of formula, it is characterized in that using the corresponding relation between paramagnetic CoAlO nano-probes and anaerobic bacteria as tie, determining in sample Anaerobic bacteria clump count.So as to determine whether the bacteriostasis of Chinese medicine is effective indirectly.Relative to anaerobic bacteria culture 2-3 days even several days Time, magnetic resonance detection only needs a few minutes.Therefore, the screening of extensive Chinese medicine and formula can be made of this method.
Embodiment
Example 1
Screening is to Propiobacterium(Anaerobic bacteria)There is the Chinese medicine preparation of suppression or therapeutic action.
1)It is prepared by nano Co AlO immunological probes:
CoAlO nano-particles can commercially, can also bibliography with either physically or chemically synthesizing.
Silicon dioxide coated nano Co AlO:47.5g sodium metasilicate is taken, is dissolved in deionized water in beaker, is adjusted with hydrochloric acid PH value is 12-13.5.0g nano Cos AlO is taken to be added in this beaker, mechanical agitation (using glass bar) 5min.By mixed liquor ultrasound 30min, in good time stirring.85 C are warming up to, salt acid for adjusting pH value 6-7, generation precipitation is added dropwise.The deionization in Magneto separate Water washing is precipitated, and is washed 3-4 times.Then, precipitation is scattered in 250mL methanol.Above procedure is in triplicate, it is ensured that silicon is depended on On CoAlO.
Amino containing silane CoAlO nano materials:Obtained silicon dioxide coated nano Co AlO is added to 25mL methanol In, use 1mLH2O and methanol dilution are to 150mL.Then the mixing of 150mL glycerine is added.Ultrasonic 30min, has been transferred to agitating device 500mL three-necked flasks in.Add 10mL amino silicane coupling agents(AEAPS), under 80-90 C after quick stirring 3h, transfer Go out product.Product is washed with deionized 3 times, and methanol washs 2 times (using Buchner funnel suction filtration).Vacuum drying.It should be noted It is that suction filtration is due to there is glycerine, so slow, supernatant liquid can be sucked with suction pipe in good time by taking out after a period of time, suction filtration process is about Need 6-8h.Obtained Aminosilylation CoAlO nano materials are finally dried in vacuo 12h.
Antibody modification:A certain amount of amido modified CoAlO nano-particles are weighed, the EDC and NHS of 2 times of quality is added, plus Enter 0.01M each 1mL of PBS.It is placed on blending instrument, 15 turns of rotating speed, reacts 45min.)The sample that soak time is arrived is carried out 10000r/min is centrifuged, time 5min, and the PBS for then adding 0.01M carries out washing centrifugation, is repeated 3 times.By the sample centrifuged Product respectively add excessive Propiobacterium antibody, and 1h is coupled on blending instrument.The 10%BSA confining liquids 45min added after the completion of coupling. After closing terminates, rotating speed 8000r/min centrifuges 5min, is centrifuged, is repeated 3 times with 0.01M PBS, then will be unnecessary Antibody, BSA are washed away, and are eventually adding PBS and are resuspended.The CoAlO nano-particles of the resistance modification of preparation are immunological probe, are protected It is stand-by in the presence of 4 DEG C.
2)The culture medium sterilized in right amount is each added in a collection of polystyrene sample pipe, acne bar is inoculated with oxygen free condition Bacterium, is separately added into after the Chinese medicine extract sealing for needing to screen, after culture a period of time, adds step 1)Prepared CoAlO Nano-probe, is sufficiently mixed concussion reaction, if there is object bacteria in sample cell, first can be anti-with CoAlO nano-probes surface Body is combined, so that CoAlO nano-probes are stayed in a liquid;If not there is object bacteria in sample cell, CoAlO nano-probes The antibody on surface can be adsorbed gradually in polystyrene sample pipe surface.After reaction 1 hour, NMR relaxation is carried out to sample cell The measure of time(Shanghai knob steps NMR20 NMRs).Using the sterile medium liquid without vaccination target bacterium as blank, The relaxation time spin-lattice relaxation time of the suspension measured has been compared with spin spin relaxation time and significantly reduced, then is said It is bright to illustrate there is object bacteria in sample cell containing probe, it is on the contrary then anti-so that the bacteriostasis of side light Chinese medicine is not obvious.Visit The amount of pin and the decline of spin-lattice relaxation time and spin spin relaxation time are proportional, by the quantitative probe of mark-on Connect the amount for quantifying object bacteria.Illustrate that the Chinese medicine added does not have inhibitory action with the presence of object bacteria.It is on the contrary then anti-.
Embodiment
Example 2
Screening is to clostridium(Anaerobic bacteria)There is the Chinese medicine preparation of suppression or therapeutic action.
1)It is prepared by nano Co AlO immunological probes:
CoAlO nano-particles bibliography physics and can also commercially can be chemically synthesized.
Silicon dioxide coated nano Co AlO:47.5g sodium metasilicate is taken, is dissolved in deionized water in beaker, is adjusted with hydrochloric acid PH value is 12-13.5.0g nano Cos AlO is taken to be added in this beaker, mechanical agitation (using glass bar) 5min.By mixed liquor ultrasound 30min, in good time stirring.85 C are warming up to, salt acid for adjusting pH value 6-7, generation precipitation is added dropwise.The deionization in Magneto separate Water washing is precipitated, and is washed 3-4 times.Then, precipitation is scattered in 250mL methanol.Above procedure is in triplicate, it is ensured that silicon is depended on On CoAlO.
Amino containing silane CoAlO nano materials:Obtained silicon dioxide coated nano Co AlO is added to 25mL methanol In, use 1mLH2O and methanol dilution are to 150mL.Then the mixing of 150mL glycerine is added.Ultrasonic 30min, has been transferred to agitating device 500mL three-necked flasks in.Add 10mL amino silicane coupling agents(AEAPS), under 80-90 C after quick stirring 3h, transfer Go out product.Product is washed with deionized 3 times, and methanol washs 2 times (using Buchner funnel suction filtration).Vacuum drying.It should be noted It is that suction filtration is due to there is glycerine, so slow, supernatant liquid can be sucked with suction pipe in good time by taking out after a period of time, suction filtration process is about Need 6-8h.Obtained Aminosilylation CoAlO nano materials are finally dried in vacuo 12h.
Antibody modification:A certain amount of amido modified CoAlO nano-particles are weighed, the EDC and NHS of 2 times of quality is added, plus Enter 0.01M each 1mL of PBS.It is placed on blending instrument, 15 turns of rotating speed, reacts 45min.)The sample that soak time is arrived is carried out 10000r/min is centrifuged, time 5min, and the PBS for then adding 0.01M carries out washing centrifugation, is repeated 3 times.By the sample centrifuged Product respectively add excessive clostridium antibody, and 1h is coupled on blending instrument.The 10%BSA confining liquids added after the completion of coupling 45min.After closing terminates, rotating speed 8000r/min centrifuges 5min, is centrifuged, is repeated 3 times with 0.01M PBS, then will Unnecessary antibody, BSA are washed away, and are eventually adding PBS and are resuspended.The CoAlO nano-particles of the resistance modification of preparation are immune Probe, be stored in 4 DEG C it is stand-by.
2)The culture medium sterilized in right amount is each added in a collection of polystyrene sample pipe, fusiform bud is inoculated with oxygen free condition Spore bacillus, is separately added into after the Chinese medicine extract sealing for needing to screen, after culture a period of time, adds step 1)Prepared CoAlO nano-probes, are sufficiently mixed concussion reaction, if there is object bacteria in sample cell, first can be with CoAlO nano-probes surface Antibody binding so that CoAlO nano-probes are stayed in a liquid.If not there is object bacteria in sample cell, CoAlO nanometers The antibody of detecting probe surface can be adsorbed gradually in polystyrene sample pipe surface.After reaction 1 hour, nuclear magnetic resonance is carried out to sample cell The measure in relaxation time(Shanghai knob steps NMR20 NMRs).Using the sterile medium liquid without vaccination target bacterium as Blank, the spin-lattice relaxation time of the suspension measured has been compared with spin spin relaxation time and significantly reduced, then is illustrated Containing probe, illustrate there is object bacteria in sample cell, it is on the contrary then anti-so that the bacteriostasis of side light Chinese medicine is not obvious.Probe Amount and the decline of spin-lattice relaxation time and spin spin relaxation time it is proportional, it is indirect by the quantitative probe of mark-on Quantify the amount of object bacteria.Illustrate that the Chinese medicine added does not have inhibitory action with the presence of object bacteria.It is on the contrary then anti-.

Claims (3)

1. a kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes, its characterization step is as follows:
1)Target bacteria antibody is coated with the preparation of paramagnetic CoAlO nano-probes;
2)The culture medium of the anaerobic bacteria of sterilizing is added in polystyrene sample pipe, vaccination target anaerobic bacteria, addition needs screening Chinese medicine preparation extract solution, after closed culture a period of time, step 1 is quantitatively adding with syringe)Prepared CoAlO nanometers are visited Pin, is sufficiently mixed concussion reaction for a period of time, if there is object bacteria in sample cell, and the antibody on CoAlO nano-probes surface is first It can be combined with object bacteria, so that CoAlO nano-probes are stayed in a liquid;If not having object bacteria in sample cell, CoAlO receives The antibody of rice detecting probe surface can be adsorbed gradually leaves nutrient solution in polystyrene sample pipe surface;Now, sample cell is carried out The measure of NMR relaxation time;Using the sterile medium liquid without vaccination target bacterium as blank, the suspension measured Relaxation time compared to significantly reducing, then explanation contains probe, so as to there is object bacteria in indirect proof sample, the amount of probe with The decline in relaxation time is proportional, and by mark-on quantitative probe, indirect quantification goes out the amount of object bacteria;There is no target in sample cell Bacterium illustrates that the traditional Chinese medicine ingredients added have a fungistatic effect, on the contrary then anti-.
2. the method according to claim 1 that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes, it is characterized in that The nano Co AlO nano-probes are the nano Co AlO materials with paramagnetic properties, and nanometer particle size is less than 300 nanometers.
3. the method according to claim 1 that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes, it is characterized in that The change in final relaxation time of the detection evaluation method based on nuclear magnetic resonance technique of described object bacteria.
CN201710200617.1A 2017-03-30 2017-03-30 A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes Pending CN107037204A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710200617.1A CN107037204A (en) 2017-03-30 2017-03-30 A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710200617.1A CN107037204A (en) 2017-03-30 2017-03-30 A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes

Publications (1)

Publication Number Publication Date
CN107037204A true CN107037204A (en) 2017-08-11

Family

ID=59534672

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710200617.1A Pending CN107037204A (en) 2017-03-30 2017-03-30 A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes

Country Status (1)

Country Link
CN (1) CN107037204A (en)

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
申静等: "基于NMR纳米探针的生物传感器用于快速检测沙门氏菌", 《食品工业科技》 *
顾立刚: "《医学免疫学与微生物学》", 31 August 2004, 中国中医药出版社 *

Similar Documents

Publication Publication Date Title
CN103217448B (en) A kind of NMR food-borne pathogen rapid detection based on paramagnetic nano Fe probe
CN107085102A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Fe Nb Cu B nano-probes
CN106841271A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on ZnO nano probe
CN107037204A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoAlO nano-probes
CN107044991A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoFeB nano-probes
CN107014851A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on FeZrB nano-probes
CN106996943A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on NdFeB nano-probes
CN107014850A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni Co nano-probes
CN107037201A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on KMnF4 nano-probes
CN106932427A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on FeAlO nano-probes
CN106841547A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoZrB nano-probes
CN106885818A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoZrO nano-probes
CN106918613A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on FeZrO nano-probes
CN107037068A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoZr nano-probes
CN106885820A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Co Cr nano-probes
CN107014848A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on FeRh nano-probes
CN107064200A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on FeSi nano-probes
CN106885817A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Fe Ni nano-probes
CN107037069A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CoPt nano-probes
CN106885821A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Cu Ni nano-probes
CN106918616A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Fe Co nano-probes
CN106885819A (en) One kind is based on NaYF4The method that nano-probe screens anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs
CN107014847A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on NiPt nano-probes
CN107044990A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on CdTe nano-probes
CN107014852A (en) A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on FeSiCo nano-probes

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20170811