CN107167469A - A kind of instruction material for differentiating gram-positive bacterium and negative bacteria and its preparation method and application - Google Patents
A kind of instruction material for differentiating gram-positive bacterium and negative bacteria and its preparation method and application Download PDFInfo
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- CN107167469A CN107167469A CN201710373459.XA CN201710373459A CN107167469A CN 107167469 A CN107167469 A CN 107167469A CN 201710373459 A CN201710373459 A CN 201710373459A CN 107167469 A CN107167469 A CN 107167469A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
Abstract
The invention discloses a kind of instruction material for differentiating gram-positive bacterium and negative bacteria, this instruction material is vancomycin and polyethyleneglycol modified golden nanometer particle, being capable of colorimetric method Rapid identification gram-positive bacterium and negative bacteria by visual observation.The gram-positive bacterium of the present invention and the instruction material non-toxic of negative bacteria, stably, preparation method it is easy, identify that bacterium is quick, need not use microscope multiplying arrangement, the deficiency of traditional dyeing identification method time length is overcome, is conducive to the quick diagnosis of bacterial disease.The invention also discloses the application that the preparation method and visual colorimetric determination of this identification gram-positive bacterium and the instruction material of negative bacteria differentiate gram-positive bacterium and negative bacteria, it will be had a good application prospect in terms of biomedical inspection, Pseudomonas discriminating, food security, environmental monitoring.
Description
Technical field
The present invention relates to a kind of method for indicating material and discriminating gram-bacteria for differentiating gram-bacteria, belong to medical science
Inspection, Pseudomonas authentication technique field.
Background technology
Bacterial cell was not only small but also transparent, and typically first to dye could be differentiated by micro- sem observation.Various dyeing
In method, the Gram's stain of Danish doctor C.Gram inventions in 1884 is a kind of most important, most widely used decoration method.
Gram's stain will contaminate at the beginning of various bacterium crystal violets first, then be decolourized with the processing of mordant iodine solution, alcohol, finally use sarranine
Redyed Deng orchil.Bacterium can be distinguished into by dyeing by two classes：Gram-positive bacterium (the Gram positive of purple
bacterium,G+)；Red gramnegative bacterium (Gram negative bacterium, G-).The difference of dyeing is main
It is due to caused by the negative difference with positive bacteria ceiis wall.After crystallized purple just dye and iodine solution mordant dyeing, in the thin of bacterium
Water insoluble crystal violet-Surgidine is formd in cell wall, gram-positive bacteria is because cell membrane is thicker, peptide glycan level of net
Many and crosslinking densification, when with ethanol or acetone decolorization, cell membrane reduces mesh because of dehydration, adds it without lipoid,
Therefore Ethanol Treatment is not in gap, therefore crystal violet and Surgidine can be firmly stayed in wall, it is still in purple to make it；And remove from office
The cell membranes of Lan Shi negative bacteriums is thin, theca externa lipid levels are high, peptidoglycan layer is thin and the degree of cross linking is poor, after decolorising agent is met, with class
Outer membrane based on fat dissolves rapidly, and thin and loose peptide glycan net can not stop the dissolution of crystal violet-Surgidine, therefore through second
Alcohol is still in colourless after decolourizing, then is redyed through orchil, just Gram-negative bacteria is taken on a red color.According to the Gram's staining of bacterium
Property, can reduce the identification scope of bacterium, be conducive to further separation identification bacterium, so as to be made to bacterial disease
Diagnosis.Because the antimicrobial spectrum of various antibiotic is different, Gram's stain is also the important references from antibiotic.
Gram's stain is widely used, but to use dyestuff iterative staining, and relies on microscope manual observation to differentiate
Bacterial species, it is cumbersome, differentiate that the time is long, it is impossible to meet the requirement of quick bacteria discriminating.Various improvement and new gram
The method of yin and yang attribute discrimination of bacteria has been reported.China Patent Publication No. CN101636415B, Martin Na Beixin in 2015
Lattice, Stefan Miller, peace merchant Philips, mikey so thatch, for being enriched with, removing and detect gram-positive bacterium
Ways and means, discloses a kind of with polypeptid specificity combination gram-positive bacteria, the method for detecting gram-positive bacteria；China
Patent publication No. CN102080121A, 2011, Edward Ruiz Lei Sendide Karstlos were used in bacteriology
Colouring method and device, disclose a kind of method and apparatus that can standardize traditional Gram's stain；Chinese patent
Publication number CN1687459, Zhang Zhiwei, Zhu Lingxiang, Wang Can, Yang Weihua, Zhang Qiong, Cheng Jing, gram-positive bacterium kind in 2005
Identification and drug resistance gene detection method and its dedicated kit, disclose a kind of amplification Gram positive bacteria strain to be measured
The primer pair and detection probe of species specificity and drug resistance gene；
Utilize porous silicon dioxide nano particle recognition gram-positive bacterium (ACS Applied Materials
Interfaces,2013,5,10874-10881)；Utilize magnetic fluorescent nanometer particle identification gram-positive bacterium (ACS
Nano,2011,5,8834-8841)；Utilize fluorescence gold nanoclusters identification gram-positive bacterium (Analytical
Chemistry, 2016,88,820-825) and utilize carbon quantum dot to recognize gram-positive bacterium (Biosensors and
Bioelectronics, 2015,74,546-553) also have been reported that.Although existing technology has perhaps to traditional dyeing identification method
Improve, it is still desirable to longer qualification time and various instrument and equipments, the discriminating to bacterium awaits exploitation faster more
Speed, easy method.
The content of the invention
Goal of the invention：The invention solves the problems that first technical problem be to provide and a kind of differentiate gram-positive bacterium and leather
The instruction material of gram-negative bacteria, this instruction material can quickly and easily differentiate Gram-positive/negative bacteria.
The invention solves the problems that second technical problem be to provide and a kind of differentiate gram-positive bacterium and Gram-negative
Bacterium indicates the preparation method of material.
The invention solves the problems that the 3rd technical problem be to provide a kind of gram-negative positive bacteria and indicate that material is differentiating
Application in terms of Gram-positive/negative bacteria.
Technical scheme：To solve above-mentioned first technical problem, the present invention provide it is a kind of differentiate gram-positive bacterium and
The instruction material of gramnegative bacterium, by the use of vancomycin and polyethylene glycol as the protective agent of golden nanometer particle, obtains ten thousand
Ancient mycin and polyethyleneglycol modified golden nanometer particle, discriminating is used as with vancomycin and polyethyleneglycol modified golden nanometer particle
Gram-positive bacterium and the instruction material of gramnegative bacterium.It is mould through the ages on this instruction material use golden nanometer particle
Element optionally combines the cell membrane of gram-positive bacterium without combining gramnegative bacterium, shows gram-positive bacterium
Red, makes gramnegative bacterium not develop the color, so as to differentiate gram-positive bacterium and gramnegative bacterium.This instruction material
Polyethylene glycol is particularly stable because having modified for material, there is long service life.
To solve above-mentioned second technical problem, the present invention provides a kind of discriminating gram-positive bacterium and Gram-negative
Bacterium indicates the preparation method of material, comprises the following specific steps that：
Step 1：A certain amount of vancomycin aqueous solution is added into certain density aqueous solution of chloraurate, is constituted mould through the ages
Element：The mol ratio of gold chloride is 0.5~20:1 mixed liquor；
Step 2：Add appropriate sodium hydroxide solution and the pH value of above-mentioned mixed liquor is adjusted to alkalescent, under stirring at room temperature instead
Answer 4 hours, add a small amount of polyethylene glycol, the lower reaction of stirring 1 hour is collected by centrifugation precipitation, centrifuged after being precipitated with milli-Q water
Separation, precipitation is resuspended in standby in ultra-pure water.
Wherein, the pH value of above-mentioned mixed liquor is adjusted to alkalescence, and alkaline pH scopes are 10-14.
Wherein, above-mentioned polyethylene glycol is the polyethylene glycol of a terminal modified sulfydryl, and molecular weight is 100-200.
To solve above-mentioned 3rd technical problem, the present invention provides a kind of discriminating gram-positive bacterium and Gram-negative
Bacterium indicates the application of material, and the application in terms of gram-bacteria is blue particular by optical colorimetry identification leather for described differentiating
Family name male/female bacterium, visual colorimetric determination identification is the red combination silver staining for the golden nanometer particle modified by vancomycin
Form what black was realized, comprise the following steps that：The solution of gram-positive bacteria/negative bacterium containing a certain amount of unknown species is led to
Cross after the method washing centrifuged, with differentiating that the suspension of instruction material of gram-positive bacterium and negative bacteria is fully mixed
Close, react 30~60 minutes at room temperature, a small amount of reacted mixed liquor is dropped in spot is formed on film, treat that spot dries on film
Afterwards, the aobvious red of spot is gram-positive bacterium, and what is do not developed the color is gramnegative bacterium；Or continue to drip on these spots
Plus silver staining reagent, after 1 minute, film is placed in hypo solution and rinsed 60 seconds, what spot showed black is gram sun
Property bacterium, what is do not developed the color is gramnegative bacterium.
Wherein, the filter paper that above-mentioned film is cellulose acetate film or composition is cellulose.
Wherein, above-mentioned silver staining reagent is made up of quinhydrones, silver nitrate and citrate or acetate.
Beneficial effect：Compared with the gram-bacteria dyeing authentication method or device reported, the present invention has following special
Color and advantage：1) the instruction material of Gram-positive/negative bacteria of the invention is due to having used the stable gold of mercapto-polyglycol
Nano-particle, stable, service life length and nontoxic, without using the organic dyestuff for easily causing distortion；2) preparation of the invention
Method is easy.Prepared by normal temperature, preparation time is short, and the experimental technique to operator is not almost required, without usual preparation method
The reaction time of middle length and heating or pyroreaction；3) method for determining bacteria of the invention is a kind of quick visual colorimetric determination identification
Method.The tedious steps such as dyestuff iterative staining, elution, heating, micro- sem observation that need not be conventional, set without by microscope amplification
It is standby；Overcome the deficiency of traditional dyeing identification method time length, be conducive to bacterial disease quick diagnosis, be adapted to field or
The occasion for lacking equipment is used.The present invention will have in terms of biomedical inspection, Pseudomonas discriminating, food security, environmental monitoring
There is good application prospect.
Brief description of the drawings
Fig. 1 differentiate that Gram-positive/negative bacteria indicates the transmission electron microscope picture of material；
Fig. 2 differentiate that Gram-positive/negative bacteria indicates the UV absorption spectrogram and color camera of material solution；
Fig. 3 differentiate that Gram-positive/negative bacteria indicates material solution and the mixed ultraviolet suction of gram-positive bacterium
Receive spectrogram；
Fig. 4 gram-positive bacteriums staphylococcus aureuses are to differentiating that Gram-positive/negative bacteria indicates the suction of material
Attached design sketch；
Fig. 5 gramnegative bacteriums Escherichia coli are to differentiating that Gram-positive/negative bacteria indicates the adsorption effect of material
Figure；
Fig. 6 gram-positive bacteriums micrococcus luteuses are to differentiating that Gram-positive/negative bacteria indicates that the absorption of material is imitated
Fruit is schemed；
Fig. 7 Gram-positive bacteria Bs bacillus is to differentiating that Gram-positive/negative bacteria indicates the absorption of material
Design sketch；
Fig. 8 differentiate that Gram-positive/negative bacteria indicates that material differentiates gram-positive bacterium and gramnegative bacterium
Result.
Embodiment
With reference to the accompanying drawings and examples, technical scheme is described in detail.
Embodiment 1 differentiates that Gram-positive/negative bacteria indicates the preparation of material
The aqueous solution of chloraurate that 5mL concentration is 1mM is added in the vancomycin aqueous solution for being 2mM to 5mL concentration, stirring is filled
Divide after mixing, the sodium hydrate aqueous solution that 0.1mL concentration is 10mM is added in mixed liquor makes the pH value of mixed liquor be 14, room temperature
The lower reaction of stirring 4 hours, by 400:1 (v/v) adds polyethylene glycol (molecular weight be 100) of the 100mg/mL containing sulfydryl, under stirring
Reaction 1 hour, golden nanometer particle is centrifugally separating to obtain by reaction solution, with milli-Q water golden nanometer particle 1 time, by Jenner's grain of rice
It is that discriminating Gram-positive/negative bacteria indicates material that son, which is resuspended in ultra-pure water and obtains cherry golden nanometer particle colloidal sol,
Material, 4 DEG C save backup.Fig. 1 is the transmission electron microscope picture of the golden nanometer particle of vancomycin modification, the gold nano of vancomycin modification
Particle has spherical shapes.Fig. 2 is the UV absorption spectrogram and color camera of the solution of gold nanoparticles of vancomycin modification, ten thousand
The solution of gold nanoparticles of ancient mycin modification is in cerise, has maximum absorption band in 520nm.
Embodiment 2 differentiates that Gram-positive/negative bacteria indicates the preparation of material
The aqueous solution of chloraurate that 5mL concentration is 1mM, stirring are added in the vancomycin aqueous solution for being 10mM to 5mL concentration
After being sufficiently mixed, the sodium hydrate aqueous solution that addition 0.1mL concentration is 10mM in mixed liquor makes the pH value of mixed liquor be 10, room
The lower reaction of temperature stirring 4 hours, by 400:1 (v/v) adds polyethylene glycol (molecular weight be 200) of the 100mg/mL containing sulfydryl, stirring
Lower reaction 1 hour, golden nanometer particle is centrifugally separating to obtain by reaction solution, with milli-Q water golden nanometer particle 1 time, by gold nano
It is that discriminating Gram-positive/negative bacteria indicates material that particle, which is resuspended in ultra-pure water and obtains cherry golden nanometer particle colloidal sol,
Material, 4 DEG C save backup.
Embodiment 3 differentiates that Gram-positive/negative bacteria indicates the preparation of material
The aqueous solution of chloraurate that 5mL concentration is 5mM is added in the vancomycin aqueous solution for being 5mM to 5mL concentration, stirring is filled
Divide after mixing, the sodium hydrate aqueous solution that 0.1mL concentration is 10mM is added in mixed liquor makes the pH value of mixed liquor be 13, room temperature
The lower reaction of stirring 4 hours, by 400:1 (v/v) adds polyethylene glycol (molecular weight be 100) of the 100mg/mL containing sulfydryl, under stirring
Reaction 1 hour, golden nanometer particle is centrifugally separating to obtain by reaction solution, with milli-Q water golden nanometer particle 1 time, by Jenner's grain of rice
It is that discriminating Gram-positive/negative bacteria indicates material that son, which is resuspended in ultra-pure water and obtains cherry golden nanometer particle colloidal sol,
Material, 4 DEG C save backup.
Embodiment 4 differentiates that Gram-positive/negative bacteria indicates the preparation of material
The aqueous solution of chloraurate that 5mL concentration is 1mM, stirring are added in the vancomycin aqueous solution for being 20mM to 5mL concentration
After being sufficiently mixed, the sodium hydrate aqueous solution that addition 0.1mL concentration is 10mM in mixed liquor makes the pH value of mixed liquor be 12, room
The lower reaction of temperature stirring 4 hours, by 400:1 (v/v) adds polyethylene glycol (molecular weight be 200) of the 100mg/mL containing sulfydryl, stirring
Lower reaction 1 hour, golden nanometer particle is centrifugally separating to obtain by reaction solution, with milli-Q water golden nanometer particle 1 time, by gold nano
It is that discriminating Gram-positive/negative bacteria indicates material that particle, which is resuspended in ultra-pure water and obtains cherry golden nanometer particle colloidal sol,
Material, 4 DEG C save backup.
Embodiment 5 differentiates that Gram-positive/negative bacteria indicates the preparation of material
The aqueous solution of chloraurate that 5mL concentration is 5mM, stirring are added in the vancomycin aqueous solution for being 2.5mM to 5mL concentration
After being sufficiently mixed, the sodium hydrate aqueous solution that addition 0.1mL concentration is 10mM in mixed liquor makes the pH value of mixed liquor be 13, room
The lower reaction of temperature stirring 4 hours, by 400:1 (v/v) adds polyethylene glycol (molecular weight be 100) of the 100mg/mL containing sulfydryl, stirring
Lower reaction 1 hour, golden nanometer particle is centrifugally separating to obtain by reaction solution, with milli-Q water golden nanometer particle 1 time, by gold nano
It is that discriminating Gram-positive/negative bacteria indicates material that particle, which is resuspended in ultra-pure water and obtains cherry golden nanometer particle colloidal sol,
Material, 4 DEG C save backup.
Embodiment 6 differentiates that Gram-positive/negative bacteria indicates selection of the material to gram-positive bacterium/negative bacteria
Property
A certain amount of gram-positive bacterium is represented into staphylococcus aureus or gramnegative bacterium represents Escherichia coli
Bacterium solution washed 3 times by the method for centrifugation, with the concentration of the quantitative bacterium solution of flow cytometer.It is 8 × 10 by concentration8, 6 ×
108, 4 × 108, 3 × 108, 2 × 108, 1 × 108, 0.5 × 108, 0 × 108Cell/mL bacterium solution 0.5mL respectively with 0.5mL
State instruction material (0.2mg/mL) mixing for differentiating gram-positive bacterium and negative bacteria that embodiment 1 is prepared, room temperature
Lower reaction 30 minutes, centrifuges supernatant and determines absorbance.
Fig. 3 is to differentiate that Gram-positive/negative bacteria indicates material and the UV absorption after staphylococcus aureus effect
Spectrogram, after differentiating that Gram-positive/negative bacteria indicates that material is mixed with staphylococcus aureus, the supernatant being centrifugally separating to obtain
Absorbance of the liquid at 520nm declines the discriminating gram sun in explanation supernatant with the increase of staphylococcus aureus concentration
Property/negative bacteria indicate material concentration decline, staphylococcus aureus concentration with the increase of staphylococcus aureus concentration
The discriminating Gram-positive of higher absorption/negative bacteria indicates that the amount of material is bigger.
Fig. 4 is the adsorption effect figure that staphylococcus aureus indicates discriminating Gram-positive/negative bacteria material, uses A0
Represent to differentiate the absorbance that Gram-positive/negative bacteria indicates material solution, A represents to differentiate Gram-positive/negative bacteria
Indicate that material centrifuges the absorbance of supernatant, staphylococcus aureus pair after being mixed with the staphylococcus aureus of various concentrations
Differentiate that Gram-positive/negative bacteria indicates that the adsorbance of material is closed with the linear direct ratio of staphylococcus aureus cell concentration
System.
Fig. 5 is gramnegative bacterium Escherichia coli to differentiating that Gram-positive/negative bacteria indicates that the absorption of material is imitated
Fruit is schemed, and Escherichia coli are to differentiating that Gram-positive/negative bacteria indicates that material is hardly combined.
Fig. 6 is gram-positive bacterium micrococcus luteus to differentiating that Gram-positive/negative bacteria indicates the absorption of material
Design sketch, gram-positive bacterium micrococcus luteus is to the adsorbance of discriminating Gram-positive/negative bacteria instruction material and its
The linear proportional relation of concentration.
Fig. 7 is Gram-positive bacteria B bacillus to differentiating that Gram-positive/negative bacteria indicates the suction of material
Attached design sketch, Gram-positive bacteria B bacillus is to differentiating that Gram-positive/negative bacteria indicates the adsorbance of material
With the linear proportional relation of its concentration.
It can be seen that from Fig. 4, Fig. 6, Fig. 7, the different gram-positive bacteriums of same amount are to differentiating that Gram-positive/feminine gender is thin
Bacterium indicates that the adsorbance of material is different.
Embodiment 7 identifies that the instruction material optical colorimetry of gram-positive bacterium and negative bacteria differentiates Gram-positive
Bacterium and gramnegative bacterium
A certain amount of gram-positive bacterium is represented into staphylococcus aureus or gramnegative bacterium represents large intestine bar
The bacterium solution of bacterium centrifuges washing 3 times, with the concentration of the quantitative bacterium solution of flow cytometer.Concentration is 80 × 109, 40 × 109, 20 ×
109, 10 × 109, 5 × 109, 2 × 109, 1 × 109What cell/mL bacterium solution 0.5mL was prepared with 0.5mL above-described embodiments 1 respectively
Differentiate instruction material (0.2mg/mL) mixing of gram-positive bacterium and negative bacteria, react 30 minutes at room temperature, centrifugation point
Separate out precipitation, then respectively plus 1mL sterilized waters form uniform suspension, 2 μ L these suspension are dropped in into cellulose acetate respectively
Spot is formed on film, after spot dries on film, the aobvious red of spot is gram-positive bacterium, and what is do not developed the color is gram-negative
Property bacterium.Or continue that 2 μ L silver staining reagents are added dropwise on these spots, after 1 minute, film is placed in 2.5% hypo solution
Middle rinsing 60 seconds, what spot showed black is gram-positive bacterium, and what is do not developed the color is gramnegative bacterium.The silver staining reagent
For quinhydrones, silver nitrate and citrate.
Fig. 8 is that the instruction material for identifying gram-positive bacterium and negative bacteria differentiates the golden yellow Portugal of gram-positive bacterium
Grape coccus (row above) and gramnegative bacterium Escherichia coli (a following row) spot figure through silver staining, microorganism concentration
It is respectively 8 × 10 from left to right10Thalline/mL, 4 × 1010Thalline/mL, 2 × 1010Thalline/mL, 1 × 1010Thalline/mL, 5 × 109
Thalline/mL, 2 × 109Thalline/mL, 1 × 109Thalline/mL, optical colorimetry differentiates the spirit of staphylococcus aureus and Escherichia coli
Sensitivity is about 5 × 109Thalline/mL.
The discriminating Gram-positive that embodiment 2~5 is prepared/negative bacteria indicates that material is obtained and embodiment 6
The identical and similar result with 7.
Claims (8)
1. a kind of instruction material for differentiating gram-positive bacterium and negative bacteria, it is characterised in that the discriminating gram sun
Property bacterium and negative bacteria indicate material be vancomycin and polyethyleneglycol modified golden nanometer particle.
2. a kind of discriminating gram-positive bacterium and negative bacteria described in claim 1 indicate the preparation method of material, it is special
Levy and be, comprise the following specific steps that：
1）A certain amount of vancomycin aqueous solution is added into certain density aqueous solution of chloraurate, vancomycin is constituted：Chlorine gold
The mol ratio of acid is 0.5 ~ 20:1 mixed liquor；
2）Add appropriate sodium hydroxide solution and the pH value of above-mentioned mixed liquor be adjusted to alkalescent, react 4 hours under stirring at room temperature,
A small amount of polyethylene glycol is added, the lower reaction of stirring 1 hour is collected by centrifugation precipitation, centrifuged after being precipitated with milli-Q water, precipitation
It is resuspended in standby in ultra-pure water.
3. according to claim 2 differentiate that gram-positive bacterium and negative bacteria indicate the preparation method of material, it is special
Levy and be, the pH value of the mixed liquor is adjusted to alkalescence, and alkaline pH scopes are 10-14.
4. according to claim 2 differentiate that gram-positive bacterium and negative bacteria indicate the preparation method of material, it is special
Levy and be, the polyethylene glycol is the polyethylene glycol of a terminal modified sulfydryl, and molecular weight is 100-200.
5. a kind of application of the instruction material of the discriminating gram-bacteria described in claim 1 in terms of gram-bacteria is differentiated.
6. application according to claim 5, it is characterised in that the application in terms of described discriminating gram-bacteria is specifically
Colorimetric method identifies Gram-positive/negative bacteria by visual observation, and visual colorimetric determination identification is the gold nano modified by vancomycin
The red of particle combines what silver staining formation black was realized, comprises the following steps that：Leather containing a certain amount of unknown species is blue
After the method washing that the solution of family name's positive bacteria/negative bacterium passes through centrifugation, with differentiating gram-positive bacterium and negative bacteria
The suspension of instruction material be sufficiently mixed, react 30-60 minute at room temperature, reacted mixed liquor on a small quantity dropped in into shape on film
Maculation, after spot dries on film, the aobvious red of spot is gram-positive bacterium, and what is do not developed the color is that Gram-negative is thin
Bacterium；Or continue that silver staining reagent is added dropwise on these spots, after 1 minute, film is placed in hypo solution and rinsed 60 seconds,
What spot showed black is gram-positive bacterium, and what is do not developed the color is gramnegative bacterium.
7. the application according to right wants 6, it is characterised in that described film is cellulose acetate film or composition is cellulose
Filter paper.
8. application according to claim 6, it is characterised in that described silver staining reagent is by quinhydrones, silver nitrate and citric acid
Salt or acetate are constituted.
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| CN111505278A (en) * | 2020-04-15 | 2020-08-07 | 西北农林科技大学 | Staphylococcus aureus detection test strip, detection method and application |
| CN113281507A (en) * | 2021-05-23 | 2021-08-20 | 吉林大学 | Rapid detection method and kit for staphylococcus aureus |
| CN113562730A (en) * | 2021-03-11 | 2021-10-29 | 海南师范大学 | Preparation of microporous gram bacteria carbon material with high specific surface area and application of microporous gram bacteria carbon material in supercapacitor |
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| CN110031438A (en) * | 2019-04-19 | 2019-07-19 | 深圳出入境检验检疫局动植物检验检疫技术中心 | A kind of activity test method of Monilinia fructicola (Winter) Honey |
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