CN107167469B - A kind of identification gram-positive bacterium and the instruction material of negative bacteria and its preparation method and application - Google Patents
A kind of identification gram-positive bacterium and the instruction material of negative bacteria and its preparation method and application Download PDFInfo
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- CN107167469B CN107167469B CN201710373459.XA CN201710373459A CN107167469B CN 107167469 B CN107167469 B CN 107167469B CN 201710373459 A CN201710373459 A CN 201710373459A CN 107167469 B CN107167469 B CN 107167469B
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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 identifying gram-positive bacterium and negative bacteria, this instruction material is vancomycin and polyethyleneglycol modified gold nanoparticle, being capable of colorimetric method Rapid identification gram-positive bacterium and negative bacteria by visual observation.Instruction material non-toxic, stabilization, the preparation method of gram-positive bacterium and negative bacteria of the invention are easy, identify that bacterium is quick, need not use microscope multiplying arrangement, the deficiency for overcoming traditional dyeing identification method time length, is conducive to the quick diagnosis of disease caused by bacterium.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 identify gram-positive bacterium and negative bacteria, will biomedical inspection, Pseudomonas identification, food safety, in terms of have a good application prospect.
Description
Technical field
The present invention relates to a kind of instruction material for identifying gram-bacteria and the methods for identifying gram-bacteria, belong to medicine
It examines, Pseudomonas authentication technique field.
Background technique
Bacterial cell was not only small but also transparent, and generally first to dye could be identified by micro- sem observation.Various dyeing
In method, the Gram's stain of invention in Danish doctor C.Gram 1884 is a kind of most important, most widely used decoration method.
Gram's stain first just contaminates various bacterium crystal violets, then is handled with mordant iodine solution, alcohol decoloration, finally uses sarranine
Equal orchils are redyed.Bacterium can be distinguished into two classes: gram-positive bacterium (the Gram positive of purple by dyeing
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
Crystal violet-Surgidine not soluble in water is formd in cell wall, gram-positive bacteria is since cell wall is thicker, peptide glycan level of net
More and crosslinking is fine and close, and when with ethyl alcohol or acetone decolorization, cell wall is because dehydration reduces mesh, in addition it is free of lipoid,
Therefore alcohol treatment is not in gap, therefore crystal violet and Surgidine can be firmly stayed in wall, keeps it still purple；And it removes from office
The cell wall of Lan Shi negative bacterium is thin, theca externa lipid levels are high, peptidoglycan layer is thin and the degree of cross linking is poor, after meeting decolorising agent, with class
Outer membrane based on rouge dissolves rapidly, and thin and loose peptide glycan net cannot stop the dissolution of crystal violet-Surgidine, therefore through second
Still it is in colourless after alcohol decoloration, then is redyed through orchil, just Gram-negative bacteria is made to take on a red color.According to the Gram's staining of bacterium
Property can reduce the identification range of bacterium, be conducive to further separate identification bacterium, to make disease caused by bacterium
Diagnosis.Since the antimicrobial spectrum of various antibiotic is different, Gram's stain is also to select the important references of antibiotic.
Gram's stain is widely used, but to use dyestuff iterative staining, and rely on microscope artificial observation to identify
Bacterial species, cumbersome, the identification time is long, is not able to satisfy the requirement of quick bacteria identification.Various improvement and new gram
The method of yin and yang attribute discrimination of bacteria has been reported.China Patent Publication No. CN101636415B, 2015, Martin Na Beixin
Lattice, Stefan Miller, peace merchant Philips, mikey so thatch, for being enriched with, removing and detecting gram-positive bacterium
Ways and means disclose 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 Karstlo used in bacteriology
Colouring method and device, disclosing a kind of can make the standardized method and apparatus of traditional Gram's stain；Chinese patent
Publication number CN1687459,2005, Zhang Zhiwei, Zhu Lingxiang, Wang Can, Yang Weihua, Zhang Qiong, Cheng Jing, gram-positive bacterium kind
Identification and drug resistance gene detection method and its dedicated kit disclose and a kind of expand 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)；Gram-positive bacterium (ACS is identified using magnetic fluorescent nanometer particle
Nano,2011,5,8834-8841)；Gram-positive bacterium (Analytical is identified using fluorescence gold nanoclusters
Chemistry, 2016,88,820-825) and carbon quantum dot is utilized to identify gram-positive bacterium (Biosensors and
Bioelectronics, 2015,74,546-553) also have been reported that.Although existing technology has perhaps traditional dyeing identification method
It is improve more, it is still desirable to longer qualification time and various instrument and equipments, to the identification of bacterium up for developing faster
Speed, easy method.
Summary of the invention
Goal of the invention: the invention solves first technical problem be to provide a kind of identification gram-positive bacterium and leather
The instruction material of gram-negative bacteria, this instruction material can quickly and easily identify Gram-positive/negative bacteria.
The invention solves second technical problem be to provide a kind of identification gram-positive bacterium and Gram-negative
The preparation method of bacterium instruction material.
The invention solves third technical problem be to provide a kind of gram-negative positive bacteria instruction material and identifying
Application in terms of Gram-positive/negative bacteria.
Technical solution: in order to solve the first technical problem mentioned above, the present invention provide a kind of identification gram-positive bacterium and
The instruction material of gramnegative bacterium obtains ten thousand using vancomycin and polyethylene glycol as the protective agent of gold nanoparticle
Ancient mycin and polyethyleneglycol modified gold nanoparticle use vancomycin and polyethyleneglycol modified gold nanoparticle as identification
The instruction material of gram-positive bacterium and gramnegative bacterium.It is mould through the ages on this instruction material use gold nanoparticle
Element keeps gram-positive bacterium aobvious selectively in conjunction with the cell wall of gram-positive bacterium without combining gramnegative bacterium
Red makes gramnegative bacterium not develop the color, to identify 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 identification 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 being added into certain density aqueous solution of chloraurate, constitutes mould through the ages
Element: the molar ratio of gold chloride is the mixed liquor of 0.5~20:1；
Step 2: appropriate sodium hydroxide solution is added by the pH value of above-mentioned mixed liquor and is adjusted to alkalescent, under stirring at room temperature instead
It answers 4 hours, a small amount of polyethylene glycol is added, stir lower reaction 1 hour, precipitating is collected by centrifugation, is centrifuged after being precipitated with milli-Q water
Separation, precipitating are resuspended in spare in ultrapure water.
Wherein, the pH value of above-mentioned mixed liquor is adjusted to alkalinity, and alkaline pH range is 10-14.
Wherein, above-mentioned polyethylene glycol is the polyethylene glycol of a terminal modified sulfydryl, molecular weight 100-200.
To solve above-mentioned third technical problem, the present invention provides a kind of identification gram-positive bacterium and Gram-negative
Bacterium indicates that the application of material, the application in terms of the identification gram-bacteria identify that leather is blue particular by optical colorimetry
Family name male/female bacterium, visual colorimetric determination identification are the red combination silver stainings for the gold nanoparticle modified by vancomycin
Form what black was realized, the specific steps are as follows: lead to gram-positive bacteria/negative bacterium solution containing a certain amount of unknown type
It is sufficiently mixed with the suspension for the instruction material for identifying gram-positive bacterium and negative bacteria after crossing the method washing of centrifuge separation
It closes, reacts 30~60 minutes at room temperature, the mixing drop after a small amount of reaction is formed into spot on film, is dried to spot 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
Add silver staining reagent, after 1 minute, film is placed in hypo solution and is rinsed 60 seconds, what spot showed black is gram sun
Property bacterium, what is do not developed the color is gramnegative bacterium.
Wherein, above-mentioned film is the filter paper that cellulose acetate film or group become cellulose.
Wherein, above-mentioned silver staining reagent is made of quinhydrones, silver nitrate and citrate or acetate.
The utility model has the advantages that the present invention has following special compared with reported gram-bacteria dyes identification method or device
Color and advantage: 1) Gram-positive/negative bacteria instruction material of the invention is due to having used mercapto-polyglycol to stablize gold
Nanoparticle is stablized, long service life and nontoxic, without using the organic dyestuff for easily causing distortion；2) preparation of the invention
Method is easy.Room temperature preparation, preparation time is short, requires the experimental technique of operator, 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.Without tedious steps such as conventional dyestuff iterative staining, elution, heating, micro- sem observations, set without by microscope amplification
It is standby；Overcome traditional dyeing identification method time long deficiency, be conducive to disease quick diagnosis caused by bacterium, be suitble to field or
The occasion for lacking equipment uses.The present invention will biomedical inspections, Pseudomonas identification, food safety, in terms of tool
There is good application prospect.
Detailed description of the invention
Fig. 1 identifies Gram-positive/negative bacteria instruction material transmission electron microscope picture；
Fig. 2 identifies Gram-positive/negative bacteria instruction material solution UV absorption spectrogram and color camera；
Fig. 3 identifies Gram-positive/negative bacteria instruction material solution and the mixed ultraviolet suction of gram-positive bacterium
Receive spectrogram；
Fig. 4 gram-positive bacterium staphylococcus aureus indicates Gram-positive/negative bacteria is identified the suction of material
Attached effect picture；
Fig. 5 gramnegative bacterium Escherichia coli indicate Gram-positive/negative bacteria is identified the adsorption effect of material
Figure；
Fig. 6 gram-positive bacterium micrococcus luteus is imitated to the absorption of Gram-positive/negative bacteria instruction material is identified
Fruit figure；
Fig. 7 Gram-positive bacteria B bacillus indicates Gram-positive/negative bacteria is identified the absorption of material
Effect picture；
Fig. 8 identifies Gram-positive/negative bacteria instruction material and identifies gram-positive bacterium and gramnegative bacterium
Result.
Specific embodiment
With reference to the accompanying drawings and examples, technical solution of the present invention is described in detail.
Embodiment 1 identifies the preparation of Gram-positive/negative bacteria instruction material
The aqueous solution of chloraurate that 5mL concentration is 1mM is added into the vancomycin aqueous solution that 5mL concentration is 2mM, stirring is filled
After dividing mixing, the sodium hydrate aqueous solution that 0.1mL concentration is 10mM, which is added, in mixed liquor makes the pH value 14 of mixed liquor, room temperature
Stirring lower reaction 4 hours, polyethylene glycol (molecular weight 100) of the 100mg/mL containing sulfydryl is added by 400:1 (v/v), under stirring
Reaction 1 hour, is centrifugally separating to obtain gold nanoparticle for reaction solution, with milli-Q water gold nanoparticle 1 time, by Jenner's grain of rice
It is to identify Gram-positive/negative bacteria instruction material that son, which is resuspended in ultrapure water and obtains cherry gold nanoparticle colloidal sol,
Material, 4 DEG C save backup.Fig. 1 is the transmission electron microscope picture of the gold nanoparticle of vancomycin modification, the gold nano of vancomycin modification
Particle has spherical shapes.Fig. 2 be vancomycin modification solution of gold nanoparticles UV absorption spectrogram and color camera, ten thousand
The solution of gold nanoparticles of ancient mycin modification is in cerise, has maximum absorption band in 520nm.
Embodiment 2 identifies the preparation of Gram-positive/negative bacteria instruction material
The aqueous solution of chloraurate that 5mL concentration is 1mM, stirring are added into the vancomycin aqueous solution that 5mL concentration is 10mM
After being sufficiently mixed, the sodium hydrate aqueous solution that 0.1mL concentration is 10mM, which is added, in mixed liquor makes the pH value 10 of mixed liquor, room
Temperature stirring lower reaction 4 hours, polyethylene glycol (molecular weight 200) of the 100mg/mL containing sulfydryl, stirring is added by 400:1 (v/v)
Lower reaction 1 hour, is centrifugally separating to obtain gold nanoparticle for reaction solution, with milli-Q water gold nanoparticle 1 time, by gold nano
It is to identify Gram-positive/negative bacteria instruction material that particle, which is resuspended in ultrapure water and obtains cherry gold nanoparticle colloidal sol,
Material, 4 DEG C save backup.
Embodiment 3 identifies the preparation of Gram-positive/negative bacteria instruction material
The aqueous solution of chloraurate that 5mL concentration is 5mM is added into the vancomycin aqueous solution that 5mL concentration is 5mM, stirring is filled
After dividing mixing, the sodium hydrate aqueous solution that 0.1mL concentration is 10mM, which is added, in mixed liquor makes the pH value 13 of mixed liquor, room temperature
Stirring lower reaction 4 hours, polyethylene glycol (molecular weight 100) of the 100mg/mL containing sulfydryl is added by 400:1 (v/v), under stirring
Reaction 1 hour, is centrifugally separating to obtain gold nanoparticle for reaction solution, with milli-Q water gold nanoparticle 1 time, by Jenner's grain of rice
It is to identify Gram-positive/negative bacteria instruction material that son, which is resuspended in ultrapure water and obtains cherry gold nanoparticle colloidal sol,
Material, 4 DEG C save backup.
Embodiment 4 identifies the preparation of Gram-positive/negative bacteria instruction material
The aqueous solution of chloraurate that 5mL concentration is 1mM, stirring are added into the vancomycin aqueous solution that 5mL concentration is 20mM
After being sufficiently mixed, the sodium hydrate aqueous solution that 0.1mL concentration is 10mM, which is added, in mixed liquor makes the pH value 12 of mixed liquor, room
Temperature stirring lower reaction 4 hours, polyethylene glycol (molecular weight 200) of the 100mg/mL containing sulfydryl, stirring is added by 400:1 (v/v)
Lower reaction 1 hour, is centrifugally separating to obtain gold nanoparticle for reaction solution, with milli-Q water gold nanoparticle 1 time, by gold nano
It is to identify Gram-positive/negative bacteria instruction material that particle, which is resuspended in ultrapure water and obtains cherry gold nanoparticle colloidal sol,
Material, 4 DEG C save backup.
Embodiment 5 identifies the preparation of Gram-positive/negative bacteria instruction material
The aqueous solution of chloraurate that 5mL concentration is 5mM, stirring are added into the vancomycin aqueous solution that 5mL concentration is 2.5mM
After being sufficiently mixed, the sodium hydrate aqueous solution that 0.1mL concentration is 10mM, which is added, in mixed liquor makes the pH value 13 of mixed liquor, room
Temperature stirring lower reaction 4 hours, polyethylene glycol (molecular weight 100) of the 100mg/mL containing sulfydryl, stirring is added by 400:1 (v/v)
Lower reaction 1 hour, is centrifugally separating to obtain gold nanoparticle for reaction solution, with milli-Q water gold nanoparticle 1 time, by gold nano
It is to identify Gram-positive/negative bacteria instruction material that particle, which is resuspended in ultrapure water and obtains cherry gold nanoparticle colloidal sol,
Material, 4 DEG C save backup.
Embodiment 6 identifies Gram-positive/negative bacteria instruction material to the selection of 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 by centrifuge separation method wash 3 times, the concentration of bacterium solution is quantified with 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 is respectively and on 0.5mL
State instruction material (0.2mg/mL) mixing for identifying gram-positive bacterium and negative bacteria that embodiment 1 is prepared, room temperature
Lower reaction 30 minutes, centrifuge separation supernatant measure absorbance.
Fig. 3 is the UV absorption identified after Gram-positive/negative bacteria instruction material and staphylococcus aureus effect
Spectrogram identifies the supernatant after Gram-positive/negative bacteria instruction material mixes with staphylococcus aureus, being centrifugally separating to obtain
Absorbance of the liquid at 520nm declines the identification gram sun illustrated in supernatant with the increase of staphylococcus aureus concentration
Property/negative bacteria instruction material concentration decline, staphylococcus aureus concentration with the increase of staphylococcus aureus concentration
The identification Gram-positive of higher absorption/negative bacteria instruction material amount is bigger.
Fig. 4 is staphylococcus aureus to Gram-positive/negative bacteria instruction material the adsorption effect figure is identified, and uses A0
It indicates to identify Gram-positive/negative bacteria instruction material solution absorbance, A indicates to identify Gram-positive/negative bacteria
Indicate the absorbance that supernatant is centrifugated after material is mixed with the staphylococcus aureus of various concentration, staphylococcus aureus pair
Identify Gram-positive/negative bacteria instruction material adsorbance and the linear direct ratio of staphylococcus aureus cell concentration closes
System.
Fig. 5 is that gramnegative bacterium Escherichia coli are imitated to the absorption of Gram-positive/negative bacteria instruction material is identified
Fruit figure, Escherichia coli hardly combine to Gram-positive/negative bacteria instruction material is identified.
Fig. 6 is gram-positive bacterium micrococcus luteus to the absorption of identification Gram-positive/negative bacteria instruction material
Effect picture, gram-positive bacterium micrococcus luteus is to identification Gram-positive/negative bacteria instruction material adsorbance and its
The linear proportional relation of concentration.
Fig. 7 is Gram-positive bacteria B bacillus to the suction of identification Gram-positive/negative bacteria instruction material
Attached effect picture, Gram-positive bacteria B bacillus indicate Gram-positive/negative bacteria is identified 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 thin to Gram-positive/feminine gender is identified
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 identifies 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 centrifuge separation of bacterium washing 3 times, the concentration of bacterium solution is quantified with 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 embodiment 1 respectively
Instruction material (0.2mg/mL) mixing for identifying gram-positive bacterium and negative bacteria, reacts 30 minutes, centrifugation point at room temperature
Precipitating is separated out, then respectively plus 1mL sterile water forms uniform suspension, 2 μ L these suspension are dripped respectively in cellulose acetate
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 to identify that the instruction material of gram-positive bacterium and negative bacteria identifies gram-positive bacterium golden yellow Portugal
Grape coccus (row above) and the spot figure of gramnegative bacterium Escherichia coli (a following row) through silver staining, microorganism concentration
It is from left to right respectively 8 × 1010Thallus/mL, 4 × 1010Thallus/mL, 2 × 1010Thallus/mL, 1 × 1010Thallus/mL, 5 × 109
Thallus/mL, 2 × 109Thallus/mL, 1 × 109Thallus/mL, optical colorimetry identify the spirit of staphylococcus aureus and Escherichia coli
Sensitivity is about 5 × 109Thallus/mL.
The identification Gram-positive that embodiment 2~5 is prepared/negative bacteria instruction material obtains and embodiment 6
With 7 identical and similar results.
Claims (8)
1. a kind of instruction material for identifying gram-positive bacterium and negative bacteria, which is characterized in that the identification gram sun
Property bacterium and negative bacteria instruction material be vancomycin and polyethyleneglycol modified gold nanoparticle, the vancomycin:
The synthesis molar ratio of gold chloride is 0.5 ~ 20:1, and the polyethylene glycol is the polyethylene glycol of a terminal modified sulfydryl, and molecular weight is
100-200。
2. a kind of preparation method for identifying gram-positive bacterium and negative bacteria instruction material described in claim 1, special
Sign is, comprises the following specific steps that:
1) a certain amount of vancomycin aqueous solution is added into certain density aqueous solution of chloraurate, constitutes vancomycin: chlorine gold
The molar ratio of acid is the mixed liquor of 0.5 ~ 20:1；
2) appropriate sodium hydroxide solution is added and the pH value of above-mentioned mixed liquor is adjusted to alkalescent, react 4 hours under stirring at room temperature,
A small amount of polyethylene glycol is added, stirs lower reaction 1 hour, precipitating is collected by centrifugation, is centrifugated after being precipitated with milli-Q water, precipitates
It is resuspended in spare in ultrapure water.
3. the preparation method according to claim 2 for identifying gram-positive bacterium and negative bacteria instruction material, special
Sign is that the pH value of the mixed liquor is adjusted to alkalinity, and alkaline pH range is 10-14.
4. the preparation method according to claim 2 for identifying gram-positive bacterium and negative bacteria instruction material, special
Sign is that the polyethylene glycol is the polyethylene glycol of a terminal modified sulfydryl, molecular weight 100-200.
5. a kind of application of instruction material described in claim 1 for identifying gram-bacteria in terms of identifying gram-bacteria.
6. application according to claim 5, which is characterized in that in terms of the identification gram-bacteria application be 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 combination silver staining of particle forms what black was realized, the specific steps are as follows: the leather containing a certain amount of unknown type is blue
After the method washing that family name's positive bacteria/negative bacterium solution passes through centrifuge separation, with identification gram-positive bacterium and negative bacteria
The suspension of instruction material be sufficiently mixed, react 30-60 minute at room temperature, by the mixing drop after reacting on a small quantity on film shape
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 is rinsed 60 seconds,
What spot showed black is gram-positive bacterium, and what is do not developed the color is gramnegative bacterium.
7. application according to claim 6, which is characterized in that the film is that cellulose acetate film or group become cellulose
Filter paper.
8. application according to claim 6, which is characterized in that the silver staining reagent is by quinhydrones, silver nitrate and citric acid
Salt or acetate are constituted.
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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 |
| 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 |
| 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 |
| CN113281507B (en) * | 2021-05-23 | 2022-08-16 | 吉林大学 | Rapid detection method and kit for staphylococcus aureus |
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| CN102439178A (en) * | 2009-05-20 | 2012-05-02 | 阿德万德克斯公司 | Methods for whole-cell analysis of gram-positive bacteria |
| CN106248597A (en) * | 2016-08-29 | 2016-12-21 | 南开大学 | A kind of visible detection method based on noble metal nano particles and application |
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| US7754444B2 (en) * | 2004-06-24 | 2010-07-13 | The Hong Kong University Of Science And Technology | Biofunctional magnetic nanoparticles for pathogen detection |
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| CN102439178A (en) * | 2009-05-20 | 2012-05-02 | 阿德万德克斯公司 | Methods for whole-cell analysis of gram-positive bacteria |
| CN106248597A (en) * | 2016-08-29 | 2016-12-21 | 南开大学 | A kind of visible detection method based on noble metal nano particles and application |
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