CN104726533A - Cold-water gellable medium coagulating agent, preparation method and application thereof - Google Patents
Cold-water gellable medium coagulating agent, preparation method and application thereof Download PDFInfo
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- CN104726533A CN104726533A CN201310706508.9A CN201310706508A CN104726533A CN 104726533 A CN104726533 A CN 104726533A CN 201310706508 A CN201310706508 A CN 201310706508A CN 104726533 A CN104726533 A CN 104726533A
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Abstract
The invention discloses a cold-water gellable medium coagulating agent, a preparation method and an application thereof. The medium coagulating agent comprises the following raw materials by weight: xanthan gum, carrageenan and guar gum, wherein mass ratio of xanthan gum to carrageenan 1-2: 1; and the mass ratio of a mixture of xanthan gum and carragheenan to guar gum is 1: 0.5-2.5. The medium coagulating agent has the advantages of no toxicity, transparency, stabilization, difficult decomposition, neutrality, high gel strength, good water retention performance and water absorption capability.
Description
Technical field
The present invention relates to a kind of culture medium solidifying agent can coagulated for the cold water of microorganism culturing and detection and its preparation method and application.
Background technology
Dull and stereotyped tilt-pour process is the classical way of microorganism detection, and have good accuracy and sensitivity, wherein culture medium solidifying agent takes on vital role in microbial culture, separation, qualification.Current agar is most widely used culture medium solidifying agent, but it exists following problem: (1) too exploitation causes expensive; (2) gel moisture easily volatilizees; (3) accumulation of condensed water easily causes between bacterium colony at culture dish wall and pollutes; (4) freezing easy generation crystallization or split, is unfavorable for that bacterial classification goes down to posterity and long term storage.In addition, because agar can not solidify in cold water, when preparing microbiological culture media, need first to heat, cool after just can be down flat plate and use, greatly have impact on detection efficiency.Therefore dull and stereotyped tilt-pour process has complex operation, time-consumingly to take a lot of work, to technician and the shortcoming such as appointed condition requirement is high.
Development fast, accurately, Micro biological Tests method is more and more subject to people's attention easily, probably can be divided into two large classes at present: a class utilizes modern molecular biology technique and automated analysis instrument to combine the microorganism method for quickly detecting set up, the bacterial count system set up as utilized impedance method, utilize the noclilucence fast detector of ATP luciferase, utilize the automatic identification system of the automatic bacterial of biochemical identification method, and enzyme joins fluorescence immunoassay (VIDAS analytical method) automatically, gold-marking immunity analytical procedure (GLISA), DNA probe and quantitative fluorescent PCR analyser etc., needed for this class techniques and methods, plant and instrument all costly, need special laboratory and professional technique operator, therefore, be difficult to penetration and promotion, another kind of is the microorganism testing slice method grown up on dull and stereotyped tilt-pour process basis, and this method simplifies dull and stereotyped tilt-pour process and improves.Microorganism testing slice method can gel etc. replace agar as carrier using the scraps of paper, non-woven fabrics or cold water, to be developed the color the microorganism detected in sample by microbial growth.Testing plate dry state is preserved, and small in volume, facilitates transport to carry; Do not need the preparation carrying out substratum before sample, directly inoculate sample, optimal temperature is cultivated and is counted, and avoids the preparation work that preparation mistake and sterilizing etc. are heavy; The waiting time cooling substratum is not needed yet.Thus testing plate method has more convenient, outstanding advantages more efficiently.The bacterium colony testing plate that such as Minnesota Mining and Manufacturing Company and oasis, Guangdong biochemical technology company limited produce, has all obtained and has applied widely.But the bacterium colony testing plate of 3M company needs through special vacuum-drying condition, and complex manufacturing, needs cost higher; Need to cultivate 48h in 37 DEG C of constant incubators, observe colony count, incubation time is oversize; Peptizer water suction is relatively slower, if careless manipulation can make sample liquid flow out from edge in application of sample process, thus affects result.
The culture medium carrier of testing plate has material impact to Detection results, and filter paper, non-woven fabrics and cold water can gel be three kinds of mainstream carrier.Domestic test card product is mainly with filter paper making carrier, and if Authorization Notice No. is the testing plate that patent discloses pathogenic micro-organism in a kind of food of CN2908517Y, this testing plate take filter paper as carrier.Mainly have the following disadvantages using filter paper as carrier: filter opening is excessive causes that filter paper is two-sided to be had colony growth, bacterium colony unintelligible and be difficult to accurate counting; Filter paper poor water retention property, inspection is not retentive of moisture again in cultivating, and is unfavorable for thalli growth, causes colony growthing slow; Also there is nutritive substance problem pockety in filter paper test card, sample liquid owing to not spreading rapidly, thus causes colony growth and skewness; Testing plate surface nutrient material is abundant not, the viscosity of gel and water retention property poor; In bacterium colony testing plate, bacterium can not be used for the preservation of bacterial classification and the propagation of bacterium.
Culture medium solidifying agent that cold water can coagulate (i.e. cold water can gel), because it is transparent, Stability Analysis of Structures, the rate of recovery are high and easily choose the advantages such as bacterium, makes with the over-all properties of its testing plate the being carrier testing plate that to be better than with filter paper, non-woven fabrics be carrier.The culture medium solidifying agent that application cold water can coagulate is as the carrier of substratum, use in the test piece of microorganism Quick Measuring or test slab instrument, do not need preheat process, the time detected can be shortened, direct dropping sample liquid several minutes just can solidify, and can carry out the operations such as bacterium colony of cultivating, count and transfer afterwards as use agar; And easy to carry, be applicable to outdoor operations.Publication number be CN101570730A patent discloses culture medium solidifying agent that a kind of cold water can coagulate and preparation method thereof.This culture medium solidifying agent main raw material is xanthan gum, guar gum, sodium polyacrylate and locust tree fresh kidney beans, but its peptizer intensity, water absorbing properties and water retention property etc. need to be improved further.
Summary of the invention
The problems such as the heat fused complex operation, the detection efficiency that cause insoluble with cold water is low are needed before agar in dull and stereotyped tilt-pour process uses in order to solve, and the problem that the peptizer intensity of the culture medium solidifying agent that can coagulate of existing cold water, water absorbing properties and water retention property have much room for improvement, the culture medium solidifying agent that main purpose of the present invention does not need before being to provide a kind of use that heat fused, cold water solubles, gel-strength are high, water absorbing properties and the excellent cold water of water retention property can coagulate.
The culture medium solidifying agent that this cold water can coagulate is made up of the raw material of following weight part: xanthan gum, carrageenin and guar gum, wherein,
The mass ratio of described xanthan gum and carrageenin is 1-2:1;
The mixture of described xanthan gum and carrageenin and the mass ratio of guar gum are 1:0.5-2.5.
Preferably, the mass ratio of described xanthan gum and carrageenin is 1:1; The mixture of described xanthan gum and carrageenin and the mass ratio of guar gum are 1:1.5.
Another object of the present invention is to provide the preparation method of the culture medium solidifying agent that above-mentioned cold water can coagulate, and comprises the following steps:
(1) pulverize xanthan gum, carrageenin and guar gum respectively, sieve;
(2) xanthan gum pulverized and sieved and carrageenin are mixed with the mass ratio of 1-2:1;
(3) mixture that step (2) obtains is mixed with the mass ratio of 1:0.5-2.5 with guar gum.
The present invention is with xanthan gum, carrageenin and guar gum for raw material, and screen filtration obtains the homogeneous mixture of particle, is the culture medium solidifying agent that cold water of the present invention can coagulate, and directly uses this culture medium solidifying agent a certain amount of to add water and can form gel.The culture medium solidifying agent that this cold water can coagulate is nontoxic, transparent, stable, be not easily decomposed, pH value is 7 ± 0.2, neutral; Peptizer intensity is high, is 300-550g/cm
2; Well, water regain is 130-150g/g, is applicable to microorganism separation and Culture for water retention property and water absorbing properties.
In order to solve existing microorganism Quick Measuring test piece (total number of bacterial colony testing plate) complex manufacturing, the problem such as cost is higher, incubation time is long, peptizer water suction slowly easily affects detected result sensitivity and accuracy, another object of the present invention is to provide the bacterium colony testing plate that a kind of manufacture craft is fairly simple, cost is low, detection time is short, highly sensitive, sensing range is wide.This bacterium colony testing plate comprises base plate, gel coat and upper strata lid from bottom to up, and the culture medium solidifying agent that wherein said gel coat can be coagulated by cold water of the present invention, nutrient broth medium, developer and somatomedin according to a certain percentage Homogeneous phase mixing form.
Preferably, the culture medium solidifying agent that in described gel coat, cold water can coagulate and the mass ratio of nutrient broth medium are 16-20:8, are more preferably 17:8.
Preferably, the mass ratio of the mixture that the culture medium solidifying agent can coagulated by cold water in described gel coat and nutrient broth medium form and developer is 750:1.
Preferably, described developer is TTC (TTC).
Preferably, described somatomedin comprises by NAD
+the somatomedin 1 that (cozymase) and acid hydrolyzed casein form and the growth factor-2 be made up of Sodium desoxycholate (DOC) and methyl glucose uronic acid sodium.
More preferably, NAD in described somatomedin 1
+the mass ratio of (cozymase) and acid hydrolyzed casein is 1:5; In described growth factor-2, the mass ratio of Sodium desoxycholate (DOC) and methyl glucose uronic acid sodium is 9:1.
Preferably, the mass ratio of the mixture that forms of above-mentioned culture medium solidifying agent, nutrient broth medium and the developer that can be coagulated by cold water and somatomedin 1, growth factor-2 is 50:1:0.5.
Preferably, described base plate selects polyester, polyethylene or polypropylene based material; Described upper strata lid selects polythene material.
Another object of the present invention is the preparation method providing a kind of bacterium colony testing plate, specifically comprises the steps:
(1) utilize double faced adhesive tape to carry out the bottom that the base plate of coating one deck presser sensor glue of appropriate size and upper strata cover bonding, double faced adhesive tape is sticked at base plate top;
(2) culture medium solidifying agent nutrient broth medium, cold water can coagulated, developer and somatomedin mix according to certain ratio uniform;
(3) mixture even spread step (2) prepared is between base plate and upper strata lid;
(4) double faced adhesive tape at base plate top is opened, by base plate and the bonding of upper strata tops, pack sealing, gas depoisoning sterilizing.
Preferably, described base plate is 10cm × 7cm, and upper strata lid is 10.5cm × 7cm.
Preferably, described in be attached to base plate top double faced adhesive tape width be 0.5cm, be pasted onto the label place at base plate top.
Bacterium colony testing plate of the present invention is saved in dull and stereotyped tilt-pour process and is prepared substratum, cleans the loaded down with trivial details step such as plate, autoclaving a kind of rhyme scheme in Chinese operas serving as the prelude to a complete score for voices, simplify the working method of microorganism detection, incubation time is reduced to present 18h by the 48h of dull and stereotyped tilt-pour process, shorten detection time, substantially increase detection efficiency, reach and save time, easy to operate rapid detection effect; Culture medium solidifying agent that this bacterium colony testing plate adopts novel cold water to coagulate replaces agar, overcomes that agar storage period is short, water retention property is poor, absorb water problem slowly, improves sensitivity and the accuracy of detection; In addition, this bacterium colony testing plate makes not to be needed through vacuum lyophilization, simplifies manufacture craft, shortens Production Time, increase work efficiency, and reduces cost of manufacture; After this bacterium colony testing plate is finished using simultaneously, can direct burning disposal, reduce environmental pollution.
Accompanying drawing explanation
Fig. 1 bacterium colony testing plate structural representation.
The water retention graphic representation of Fig. 2 different culture media peptizer.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described.Envrionment conditions required in the embodiment of the present invention is temperature 27-30 DEG C, humidity 30-40%.
Xanthan gum and guar gum: Zibo Deosen Biochemical Ltd..
Carrageenin: good harvest bio tech ltd, Xi'an.
Embodiment 1: the preparation of the culture medium solidifying agent that cold water can coagulate
Preparation example 1: xanthan gum, carrageenin and guar gum after pulverizing are crossed 160 mesh sieves respectively, and getting each 5g, 5g, 15g(mass ratio of xanthan gum, carrageenin and guar gum is 1:1:3) mix, namely obtain the culture medium solidifying agent that cold water can coagulate.
Preparation example 2: xanthan gum, carrageenin and guar gum after pulverizing are crossed 160 mesh sieves respectively, and getting each 6g, 6g, 12g(mass ratio of xanthan gum, carrageenin and guar gum is 1:1:2) mix, namely obtain the culture medium solidifying agent that cold water can coagulate.
Preparation example 3: xanthan gum, carrageenin and guar gum after pulverizing are crossed 160 mesh sieves respectively, and getting each 8g, 8g, 8g(mass ratio of xanthan gum, carrageenin and guar gum is 1:1:1) mix, namely obtain the culture medium solidifying agent that cold water can coagulate.
The water retention property simultaneous test of embodiment 2 different culture media peptizer
Test method:
Get the xanthan gum after pulverizing, carrageenin, guar gum respectively, cross 160 mesh sieves, as the culture medium solidifying agent of test group (1) ~ (3);
The culture medium solidifying agent that in Example 1, the cold water of preparation example 1 ~ 3 can coagulate, as the culture medium solidifying agent of test group (4) ~ (6);
According to the content disclosed in the patent of CN101570730A of publication number in background technology part, get 5g xanthan gum and 5g guar gum (mass ratio is 1:1) mixing, by 1g polyacrylic acid and 8g Viscogum BE (mass ratio is 1:8) mixing, then above-mentioned front latter two mixture is mixed with the mass ratio of 8:1, obtain the culture medium solidifying agent that another cold water can coagulate, as the culture medium solidifying agent of test group (7);
The culture medium solidifying agent 1g weighed in above-mentioned test group (1) ~ (7) is put in the plate of diameter 7cm respectively, add 25ml(25g) deionized water, mixing, then its mass percent is 4%, after fully swelling, measure weight (ml), put into 37 DEG C of thermostat containers and cultivate and open plate lid, measure a weight (m respectively at 4h, 8h, 16h, 20h
2), press the water retention of formulae discovery different time points: water retention=m
2/ m
1.Each test group gets 5 samples, is averaging water retention.
In contrast with blank water sample, investigate the water retention property of gel simultaneously.During blank water sample 0h, measure weight (ml), put into 37 DEG C of thermostat containers cultivate and open plate lid, measure a weight (m2) respectively at 4h, 8h, 16h, 20h, press the water retention of formulae discovery different time points: water retention=m2/m1.Be the water retention of blank water sample control group.
Test-results:
Test-results is shown in Fig. 2.Visible, the water retention of the culture medium solidifying agent of test group (4) section is at one time all higher than other culture medium solidifying agent, and therefore the culture medium solidifying agent of test group (4) has good water retention property.
Embodiment 3: the water regain of the culture medium solidifying agent that cold water can coagulate and gel-strength simultaneous test
Test method: each 0.5g of culture medium solidifying agent powder that the cold water distinguishing test group (7) gained in Example 1 preparation example 1 and embodiment 2 can coagulate, adds 100m1 deionized water, 121 DEG C of sterilizing 15min in Erlenmeyer flask; Draw 10m1, drip in diameter 7cm dull and stereotyped, pH value is 7.2-7.4, and 50 DEG C of dry 7h, survey the water regain of its gel.Gel-strength is measured with BROOKFIELDLFRA-4500 gel strength measuring instrument.Use determinator optimum configurations: trigger point 3.0g, distance 0.2mm, speed 0.1mm/s, cylindrical probe area 1.266cm
2.Strength formula is: gel-strength (g/cm
2)=measure reading // 1.266.
Test-results:
The culture medium solidifying agent water regain that in embodiment 1, preparation example 1 gained cold water can coagulate is 140g/g, and pH value is 7.2, and gel-strength is 450.0g/cm
2;
The culture medium solidifying agent water regain that in embodiment 2, the cold water of test group (7) gained can coagulate is 107g/g, and pH value is 7.4, and gel-strength is 376.0g/cm
2.
Contrast visible, the water absorbing properties of the culture medium solidifying agent that cold water can coagulate in the embodiment of the present invention 1 preparation example 1 and gel-strength are all better than the publication number culture medium solidifying agent that cold water can coagulate disclosed in the patent of CN101570730A.
Embodiment 4: the preparation of bacterium colony testing plate and performance test
Bacterium colony testing plate is prepared in the culture medium solidifying agent utilizing cold water of the present invention to coagulate, and concrete steps are as follows:
1. get culture medium solidifying agent 35g, the nutrient broth medium 16.47g that can coagulate according to the cold water of the method gained of preparation example 1 in embodiment 1, TTC (TTC) 68.6mg, mix to obtain mixture (1);
2. weigh dehydrogenase coenzyme (NAD respectively according to 2:1:1:5 ratio
+) 0.25g and acid hydrolyzed casein 1.25g, mix to obtain mixture (2);
3. weigh Sodium desoxycholate (DOC) 0.45g and methyl glucose uronic acid sodium 0.05g respectively according to 9:1 ratio, mix to obtain mixture (3);
4. get said mixture (1) 51.54g, mixture (2) 1g and mixture (3) 0.5g, mix;
5. utilize double faced adhesive tape 3 to bond the bottom of the base plate 1 of coating one deck presser sensor glue of appropriate size and upper strata lid 2, double faced adhesive tape is sticked at base plate top;
6. the above-mentioned mixture finally obtained is uniformly coated between base plate 1 and upper strata lid 2;
7. open the double faced adhesive tape at base plate 1 top, the top of base plate 1 and upper strata lid 2 is bonded;
8. packing, sealing, ethylene oxide sterilizing, 4 DEG C keep in Dark Place for subsequent use.
The method for testing performance of bacterium colony testing plate:
The test method of bacterium colony testing plate water regain and gel-strength is with embodiment 3.
Test-results:
The water regain of bacterium colony testing plate is 124g/g, pH is 7.3, and gel-strength is 300g/cm
2.
Embodiment 5: the bacterium colony testing plate utilizing embodiment 4 to prepare and dull and stereotyped tilt-pour process measure the simultaneous test of total number of bacterial colony
Test method:
Select several representational bacterial classifications such as intestinal bacteria 8099, streptococcus aureus 6538, suis 556, pasteurella multocida C48-3, Salmonellas 1789, often kind of bacterial classification gets 8 samples, be inoculated in the bacterium colony testing plate of embodiment 4 preparation respectively, dilution bacterium liquid, adjustment bacterial concentration is 30<X<300, and quantity of microorganism inoculated is 1ml.In 37 DEG C of thermostat containers, cultivate 48h, calculate red colonies sum.Detect corresponding bacterial classification with the dull and stereotyped tilt-pour process of GB, adjustment bacterial concentration is 30<X<300, and quantity of microorganism inoculated is 1ml simultaneously.Then will dissolve and the suitable nutrient agar being cooled to 45 DEG C is poured in flat board, swing plate gently, makes the bacterium liquid of dilution mix with substratum, and after agar solidification, upset is dull and stereotyped, in 37 DEG C of thermostat containers, cultivate 48h, calculates total number of bacterial colony.Test-results:
Each bacterial classification comparative test result is in table 1 ~ 5.Visible, bacterium colony testing plate prepared by embodiment 4 is compared with dull and stereotyped tilt-pour process, for the measurement result difference all remarkable (P>0.05) of 5 kinds of bacterial classifications.
Table 1 E. coli clones sum
Table 2 S. aureus colonies sum
Table 3 suis total number of bacterial colony
Table 4 pasteurella multocida total number of bacterial colony
Table 5 Salmonellas total number of bacterial colony
Embodiment 6: the total number of bacterial colony in the bacterium colony testing plate testing environment utilizing embodiment 4 to prepare
Test method:
Utilize the bacterium colony testing plate that embodiment 4 makes, the total number of bacterial colony in testing environment.Respectively at 30min before sterilization, after sterilization in farming site surface sample.The sample that same operation gathers 2 plants (each plant gathers 3 animal colony houses) detects.
The concrete method of sampling is: carry out preparation disinfectant solution according to product description, carries out air-atomizing, reach sterilisation purpose with atomizer.The lattice needing the ground of sampling to be divided into three 5cm × 5cm, 30min before being labeled as sterilization, after sterilization.Being bedewed in the physiological saline of 1ml by sterile cotton swab during sampling, is 5cm × 5cm region swabing sampling to area, and during sampling, cotton swab is rotated on sampling limit in limit, anyhow comes and goes 10 times.After sampling, discard the cotton swab end of hand contact, aseptic sampling end is inserted in the cillin bottle containing 5ml physiological saline, sealing.With electronic vortex mixer concussion 30s, bacterium wash-out is obtained elutriant, then carry out 10 times (1ml elutriant+9ml physiological saline) dilution and 100 times (1ml10 times of diluent+9ml physiological saline) dilution.10 times of diluents are got respectively and 100 times of diluent 1ml bacterium colony testing plate detect before sterilization.0.5h after sterilization, cancels the former elutriant after poison and its 10 times of diluent 1ml bacterium colony testing plate detect.
Bacterium colony testing plate detection method is as follows: bacterium colony testing plate taken out from refrigerator, be placed to room temperature, open sealed bag to take out, open upper strata lid, add elutriant 1ml in central authorities, with circular matching die compressing tablet, to be solidified after 1min, put in 37 DEG C of incubators and cultivate 16-18h, observations, the sterilizing rate of sterilizing agent can be calculated, for the evaluation of sterilizing agent to pathogenic micro-organism sterilisation effect in environment.
Test-results: the total number of bacterial colony detected result in plant's environment is in table 6.
Total number of bacterial colony detected result in table 6 plant environment
Embodiment 7: in different total number of bacterial colony measuring method, incubation time affects simultaneous test to total number of bacterial colony
Test method:
Choose identical bacterial classification, the dull and stereotyped tilt-pour process of the bacterium colony testing plate utilizing embodiment 4 to prepare and the world measures the total number of bacterial colony of different time points.Specific as follows: picking intestinal bacteria 8099, streptococcus aureus 6538 lawn, be inoculated in 5ml nutrient broth medium respectively, after cultivating 18-24h, get intestinal bacteria liquid and the 1ml streptococcus aureus liquid of 1ml respectively, mix rear dilution bacterium liquid, adjustment bacterial concentration is 30<X<300, open the upper strata lid of bacterium colony testing plate prepared by embodiment 4, the bacterium liquid 1ml regulated is added in middle position, with circular matching die compressing tablet, to be solidified after 1min, put in 37 DEG C of incubators and cultivate 18h, 24h and 48h, observations is also carried out enumeration and is obtained total number of bacterial colony.Detect total number of bacterial colony with the dull and stereotyped tilt-pour process of GB simultaneously.
Detected result:
The detected result of two kinds of detection methods is in table 7.Visible, the bacterium colony testing plate adopting embodiment 4 to prepare detects the total number of bacterial colony obtained and the result there was no significant difference (P>0.05) adopting dull and stereotyped tilt-pour process to cultivate 48h when cultivating 18h, in other words, adopt bacterium colony testing plate prepared by embodiment 4, the total number of bacterial colony that incubation time detects when being 18h can reach the total number of bacterial colony that dull and stereotyped tilt-pour process cultivation 48h detects, has greatly saved detection time.
Table 7 incubation time affects test-results to total number of bacterial colony
Note: Biao “ ﹡ " represent and detect total number of bacterial colony with bacterium colony testing plate compared with, there was no significant difference, P > 0.05.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. the culture medium solidifying agent that can coagulate of cold water, is characterized in that, be made up of the raw material of following weight part: xanthan gum, carrageenin and guar gum, wherein,
The mass ratio of described xanthan gum and carrageenin is 1-2:1;
The mixture of described xanthan gum and carrageenin and the mass ratio of guar gum are 1:0.5-2.5.
2. the culture medium solidifying agent that the cold water according to claims 1 can coagulate, is characterized in that, the mass ratio of described xanthan gum and carrageenin is 1:1; The mixture of described xanthan gum and carrageenin and the mass ratio of guar gum are 1:1.5.
3. the cold water preparation method of culture medium solidifying agent that can coagulate, is characterized in that, comprise the following steps:
(1) pulverize xanthan gum, carrageenin and guar gum respectively, sieve;
(2) xanthan gum pulverized and sieved and carrageenin are mixed with the mass ratio of 1-2:1;
(3) mixture that step (2) obtains is mixed with the mass ratio of 1:0.5-2.5 with guar gum.
4. a bacterium colony testing plate, it is characterized in that, comprise base plate from bottom to up, gel coat and upper strata lid, the culture medium solidifying agent that wherein said gel coat can be coagulated by the cold water described in claim 1 or 2, nutrient broth medium, developer and somatomedin according to a certain percentage Homogeneous phase mixing form.
5. bacterium colony testing plate according to claim 4, is characterized in that, the culture medium solidifying agent that in described gel coat, cold water can coagulate and the mass ratio of nutrient broth medium are 16-20:8.
6. bacterium colony testing plate according to claim 4, is characterized in that, the mass ratio of the mixture that the described culture medium solidifying agent that can be coagulated by cold water and nutrient broth medium form and developer is 750:1.
7. bacterium colony testing plate according to claim 4, is characterized in that, described developer is TTC.
8. bacterium colony testing plate according to claim 4, is characterized in that, described somatomedin comprises by NAD
+the somatomedin 1 that (cozymase) and acid hydrolyzed casein form and the growth factor-2 be made up of Sodium desoxycholate (DOC) and methyl glucose uronic acid sodium; NAD in described somatomedin 1
+the mass ratio of (cozymase) and acid hydrolyzed casein is 1:5, and in described growth factor-2, the mass ratio of Sodium desoxycholate (DOC) and methyl glucose uronic acid sodium is 9:1.
9. bacterium colony testing plate according to claim 8, is characterized in that, can coagulate mixture that culture medium solidifying agent, nutrient broth medium and developer form and somatomedin 1, the mass ratio of growth factor-2 is 50:1:0.5 in described gel coat by cold water.
10. a preparation method for bacterium colony testing plate, specifically comprises the steps:
(1) utilize double faced adhesive tape to bond the bottom of the base plate of appropriate size and upper strata lid, double faced adhesive tape is sticked at base plate top;
(2) the culture medium solidifying agent can coagulated by cold water, nutrient broth medium, developer and somatomedin mix according to certain ratio uniform;
(3) mixture even spread step (2) prepared is between base plate and upper strata lid;
(4) double faced adhesive tape at base plate top is opened, by base plate and the bonding of upper strata tops, gas depoisoning sterilizing, pack sealing.
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| CN105754849A (en) * | 2016-03-31 | 2016-07-13 | 吉林农业大学 | Total bacterial count testing slip and preparation method and application thereof |
| CN106868092A (en) * | 2016-12-29 | 2017-06-20 | 贵州科学院 | A kind of method of rural potable water microorganism field quick detection |
| CN108220384A (en) * | 2017-12-29 | 2018-06-29 | 广东海大畜牧兽医研究院有限公司 | A kind of vibrios testing piece and preparation method thereof |
| CN112724474A (en) * | 2020-12-25 | 2021-04-30 | 深圳市航天食品分析测试中心有限公司 | Cold hydrogel and application thereof |
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| CN104911243A (en) * | 2015-06-29 | 2015-09-16 | 汇征联合（北京）医疗器械有限公司 | Solid culture medium for inoculating liquid samples and culture method |
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| CN105754849B (en) * | 2016-03-31 | 2018-02-16 | 吉林农业大学 | A kind of total plate count test piece, preparation method and applications |
| CN106868092A (en) * | 2016-12-29 | 2017-06-20 | 贵州科学院 | A kind of method of rural potable water microorganism field quick detection |
| CN108220384A (en) * | 2017-12-29 | 2018-06-29 | 广东海大畜牧兽医研究院有限公司 | A kind of vibrios testing piece and preparation method thereof |
| CN112724474A (en) * | 2020-12-25 | 2021-04-30 | 深圳市航天食品分析测试中心有限公司 | Cold hydrogel and application thereof |
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