CN106556599B - A kind of method for rapid inspecting animalcule - Google Patents
A kind of method for rapid inspecting animalcule Download PDFInfo
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- CN106556599B CN106556599B CN201610936062.2A CN201610936062A CN106556599B CN 106556599 B CN106556599 B CN 106556599B CN 201610936062 A CN201610936062 A CN 201610936062A CN 106556599 B CN106556599 B CN 106556599B
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Abstract
The invention discloses a kind of method for rapid inspecting animalcule, in specifically liq culture medium enrichment culture sample during the entire process of objective microbe, the small molecule product being metabolized when objective microbe is grown (such as carbon dioxide, small molecule acid) is detected by organosilicon material and the sensor of color indicator solution composition using a kind of, such metabolite, which penetrates into sensor, causes the color of sensor to change, and comes to whether there is objective microbe in judgement sample with this.This method can effectively shorten detection Institute of Micro-biology time, also have many advantages, such as it is of low cost, use manpower and material resources sparingly, testing result it is reliable.
Description
Technical field
The invention belongs to microorganism detection fields, and in particular to a kind of method for rapid inspecting animalcule.
Background technology
In the fields such as food, drug and Cosmetic Manufacture, health supervision, in order to ensure product quality, from adopting for raw material
Finished product manufacture is bought, has corresponding quality control standard, wherein microorganism detection is exactly conventional Quality Control project.It is most popular micro-
Biological detecting method is mainly plating method, is separately cultured the microorganism to survive in sample using special media, makes
These microorganisms to live can form the bacterium colony of various forms on tablet.This method is sensitive, intuitive, accurate, can provide simultaneously
The value volume and range of product of microorganism in test sample.But plating method has the following disadvantages：
1, time-consuming, and 1~5 day time is needed from starting to cultivate to forming macroscopic bacterium colony；
2, cumbersome, prepare tablet, culture, bacterium colony count, need during biochemical identification largely sterilize to go out
Bacterium and cleaning will expend a large amount of man power and material；Height is required to the technical merit of operating personnel, is susceptible to artificial mistake
Difference；
3, testing result lag, can not quickly, detection information is provided in real time, be unfavorable for production and superintendent office take in time
Satisfactory after correcting.Although plating method improves to some extent in recent years, but still can not obtain testing result in real time, use cannot be met
Department finds in time and control pollution, acceleration product are let pass, reduce warehousing costs, speed up capital turnover, adjustment produces pin in time
The demands such as the plan of selling.
Therefore, there is an urgent need for research and develop a kind of method for rapid inspecting animalcule.
Invention content
In order to solve the deficiencies in the prior art, the present invention provides a kind of method for rapid inspecting animalcule, this method can have
Effect shorten detection Institute of Micro-biology time, also have many advantages, such as it is of low cost, use manpower and material resources sparingly, testing result it is reliable.
In order to solve the above technical problems, the present invention provides a kind of method for rapid inspecting animalcule, in specifically liq culture
In base enrichment culture sample during the entire process of objective microbe, using one kind by organosilicon material and color indicator solution group
At sensor come the small molecule product that is metabolized when detecting objective microbe growth（Such as carbon dioxide, small molecule acid）, this
Class metabolite, which penetrates into sensor, causes the color of sensor to change, and comes to whether there is mesh in judgement sample with this
Mark microorganism.
As currently preferred technical solution, the effect of the specifically liq culture medium is carried for objective microbe
For abundant nutriment, promote growth breeding, while inhibiting the growth of other non-targeted microorganisms, the specifically liq
Culture medium using nutrient broth, pancreas peptone soybean broth, malt juice liquid medium, green lactose bile salt broth, AC meat soups,
Bu Shi-Haas meat soup, potato dextrose broth, PA meat soups, RV meat soups and it is other to various objective microbes have growth-promoting
Grow any one in the culture solution of effect.
As currently preferred technical solution, the organosilicon material includes base polymer, crosslinking agent and catalysis
Agent, wherein the base polymer refers to the organopolysiloxane containing vinyl；The crosslinking agent refers to having for silicon-hydrogen bond containing
Machine polysiloxanes；The catalyst refers to organic tin catalyst, platinum group catalyst or titanate esters and its composition catalyst；
The content of each component is：78wt%-99wt% base polymers, 0.5wt%-20wt% crosslinking agents, 0.5wt%-2wt% catalyst；
The content of color indicator is 0.01wt%-1wt% in the color indicator solution；The organosilicon material is indicated with color
The weight ratio of agent solution is 3:(0.15-0.5).
As currently preferred technical solution, the base polymer be α, ω-divinyl polydimethyl silicone oil,
Or methyl vinyl MQ silicon resin；The crosslinking agent is linear methyl hydrogen polysiloxanes, cyclic methyl hydrogen polysiloxane or methyl
Hydrogen MQ silicones；The catalyst is dihydroxy acid dialkyl tin, dicarboxylic acids stannous, platinum-vinyl siloxane complex, alcohol modification
Chloroplatinic acid catalyst, platinum-alkynes base complex or monoalkoxy type titanate esters；The color indicator is selected from bromocresol green, phenol
It is phthalein, phenol red, cresol red, reindeer moss, thymol blue, bromthymol blue, pyrocatechol purple, azolitmin, quinone quinoline imines, bright orange, neutral
Red, rosolic acid, rheochrysin, 2,2 '-dihydroxy benzenes ketenes, alphanaphthol phthalein, meta cresol purple, curcumin, xylenol blue and
It is one or more in other color indicators.
As currently preferred technical solution, the sensor is opaque or translucent elastic solid, initial face
Color is black, brown, dark brown, blackish green and other dark colours, because organosilicon material has highly-breathable and selective
Only allow certain gases and small-molecule substance to pass through, and other liquid, solid and microorganism cannot all enter, therefore can be significantly
Exclusion sample interference, the only small-molecule substance in microorganism growth metabolism product（Such as carbon dioxide, small molecule acid）
It can just penetrate into sensor and the color of sensor is caused to eventually become yellow, red, orange red or orange-yellow, and make
The light transmittance or index of refraction of sensor increase.The microbial metabolic products that the sensor detects are more, used in its discoloration
Time it is shorter, the light transmittance or index of refraction of sensor are bigger.
As currently preferred technical solution, this method includes preparing a kind of device of quick detection microorganism, the dress
It sets comprising upper layer, lower layer's two parts, specifically liq culture medium built in upper layer, lower layer's built-in sensors, sensor and specificity
Fluid nutrient medium is in direct contact.
As currently preferred technical solution, a kind of device of quick detection microorganism is cylinder, pros
The transparent resealable container of body, cuboid or other suitable shapes.
As currently preferred technical solution, including step used below：
(1) sample is handled：Sample is done into appropriate dilution with sterile diluent, fluid sample can not have to dilution yet and handle；
(2) the device in right amount through step (1) treated sample to the quick detection microorganism is pipetted
In, it is uniformly mixed after sealing；
(3) cultivate：The device of the quick detection microorganism of step (2) is placed at the suitable temperature of objective microbe and is cultivated,
The culture 1-24h of bacterium class, the culture 1-48h of Mycophyta；
(4) judging result：Whether visually observe sensor priming color in the device of the quick detection microorganism of step (3)
It changes, what priming color changed then shows do not have with the presence of objective microbe, priming color in sample
What is changed then shows do not have objective microbe presence in sample.
As the preferred technical solution of the present invention, (4) the realization means of automation may be used in the step, i.e.,：Using outer
Portion's light source and photon detection technology monitor the variation of sensor light transmittance caused by color change or diopter in real time, pass through
Computer control system, which automatically analyzes, quickly determines objective microbe content.
Compared with prior art, the present invention has following technique effect：For opposing plates cultural method of the present invention, bacterium
Detection only needs 1-24h, the detection of mould saccharomycete only to need 1-48h, hence it is evident that shortens detection time, microbial detection device is pre- in advance
System is taken and is used, the work of a large amount of configuration culture mediums, cleaning glass apparatus is omitted.If in conjunction with new light sources and photon detection skill
Art is changed using the light transmittance or index of refraction of computer control system monitoring sensor, can know testing result in real time.
Description of the drawings
Fig. 1 is a kind of structural schematic diagram of the device of quick detection microorganism in the present invention.
Specific implementation mode
With reference to specific embodiment, the present invention is furture elucidated, but these embodiments are only intended to illustrate the present invention, and
It does not limit the scope of the invention.
Embodiment 1
As shown in Figure 1, a kind of device of quick detection microorganism of the present invention, including bottle cap 1 and body 2, it is upper that body 2, which divides,
Two layer, lower layer parts：Upper layer is cultivation region 21, includes specifically liq culture medium；Lower layer is detection zone 22, built-in sensing
Device.The specifically liq culture medium can be specifically nutrient broth herein, be used for detection bacterium sum.The quick inspection
The device preparation process of micrometer biology is as follows：
1, sensor is prepared：
（1）Prepare color indicator solution：0.5wt% bromthymol blues, 0.05wt% xylenol blues are dissolved in 20% ethyl alcohol
In solution；
（2）Mix organosilicon material and color indicator solution：Weigh the linear first that hydrogen content is 0.24% successively first
Base hydrogen polysiloxanes 0.8g, α, ω-divinyl polydimethyl silicone oil 28.5g and 10ppm platinum-vinyl siloxane complex
Then 0.7g presses organosilicon material in plastic cup：Color indicator solution=3:It is molten that 0.2 weight ratio weighs color indicator
Liquid is uniformly mixed in same plastic cup, so that indicator is dispersed in organosilicon material, be sub-packed in small bottom of bottle with cover
Portion, about 1cm thickness are coagulation forming after standing 1-24 hours.
2, filling and sterilized liquid culture medium：
A. nutrient broth is poured into sensor in the bottle of coagulation forming, the volume of nutrient broth is the 2/ of bottle volume
3；
B. after covering bottle cap, bottle is put into high-pressure sterilizing pot and is sterilized by the sterilizing requirement of nutrient broth；
C. finished product can be obtained after taking out.
Using：When using this microbial detection device, sample is done into proper ratio with sterile diluent first
Then dilution takes 1mL samples to dilute even liquid and is added in the device, tighten liquid in bottle cap mixing bottle and be just placed at 36 DEG C and cultivate
1-24h.The sensor of the bottom of device turns yellow, and indicates that the sample added dilutes in even liquid with the presence of bacterium.The device is added
In sample contained by objective microbe quantity it is more, the sensor discoloration time used is shorter.
Contrast test has been carried out with number of patent application PCT/US2010/003242 described devices below for the present invention, it is main
Detection time is compared, concrete outcome is shown in Table 1.
The data comparison table of table 1 present invention and number of patent application PCT/US2010/003242 described devices
| Sample | Present invention detection duration used | PCT/US2010/003242 invention detections duration used | The clump count and duration used that the detection of GB4789.2-2010 plating methods comes |
| Bacterium solution 1 | 1.8 h | 2.0 h | 4.0×108 CFU/mL, 48h |
| Bacterium solution 2 | 2.5 h | 2.7 h | 3.2×107 CFU/mL, 48h |
| Bacterium solution 3 | 3.8 h | 4.0 h | 3.8×106CFU/mL, 48h |
| Bacterium solution 4 | 5.6 h | 5.8 h | 4.2×105 CFU/mL, 48h |
| Bacterium solution 5 | 6.9 h | 7.0 h | 3.6×104 CFU/mL, 48h |
| Bacterium solution 6 | 8.0 h | 8.3 h | 5.1×103 CFU/mL, 48h |
| Bacterium solution 7 | 10.5 h | 11.3 h | 4.5×102 CFU/mL, 48h |
| Bacterium solution 8 | 14.2 h | 15.2 h | 6.0×101 CFU/mL, 48h |
Bacterium solution in table 1 is the bacterium colony sterile saline using arbitrary picking on fresh plate count agar tablet
Manufactured 10 times of gradient dilution bacterium solutions, then take 1mL that PCT/US2010/003242 institutes are respectively added respectively from each gradient bacterium solution
Device and device of the present invention are stated, then is put into simultaneously with transmitting light source and photon detector automatic microbe culture detection system
Detection is cultivated in system, obtains the data in above-mentioned table 1.Another each bacterium solution simultaneously using GB4789.2-2010 plating methods come
Detect clump count.As shown in table 1, compared with PCT/US2010/003242 described devices, detection time of the invention is shorter,
It can be seen that present invention reduces the times of detection Institute of Micro-biology.
Embodiment 2
As shown in Figure 1, a kind of device of quick detection microorganism of the present invention, including bottle cap 1 and body 2, it is upper that body 2, which divides,
Two layer, lower layer parts：Upper layer is cultivation region 21, includes specifically liq culture medium；Lower layer is detection zone 22, built-in sensing
Device.The specifically liq culture medium can be specifically malt juice liquid medium herein, for detecting mould saccharomycete.
The device preparation process of the quick detection microorganism is as follows：
1, sensor is prepared：
（1）Prepare color indicator solution：0.2wt% bromthymol blue sodium salts, the phenol red sodium salts of 0.3wt%, are dissolved in distilled water
Middle heating for dissolving；
（2）Mix organosilicon material and color indicator solution：Weigh the linear first that hydrogen content is 0.8% successively first
Base hydrogen polysiloxanes 0.15g, α, ω-divinyl polydimethyl silicone oil 29.7g and 30ppm alcohol are modified chloroplatinic acid catalyst
Then 0.15g presses organosilicon material in plastic cup：Color indicator solution=3:It is molten that 0.3 mass ratio weighs color indicator
Liquid is uniformly mixed in same plastic cup, so that indicator is dispersed in organosilicon material, is sub-packed in bottle bottom with cover,
About 1cm thickness is coagulation forming after standing 1-24 hours.
2, filling and sterilized liquid culture medium：
A. malt juice liquid medium is poured into sensor in the bottle of coagulation forming, the body of malt juice liquid medium
Product is the 1/3 of bottle volume；
B. after covering bottle cap, bottle is put into high-pressure sterilizing pot and is sterilized by the sterilizing requirement of nutrient broth；
C. finished product can be obtained after taking out.
Using：When using this microbial detection device, sample is done into proper ratio with sterile diluent first
Then dilution takes 1mL samples to dilute even liquid and is added in the device, tighten liquid in bottle cap mixing bottle and be just placed at 28 DEG C and cultivate
1-48h.The sensor of the bottom of device turns yellow, and indicates that the sample added dilutes in even liquid with the presence of mould saccharomycete.It is added
The quantity of objective microbe contained by sample in the device is more, and the time used in sensor discoloration is shorter.
Embodiment 3
As shown in Figure 1, a kind of device of quick detection microorganism of the present invention, including bottle cap 1 and body 2, it is upper that body 2, which divides,
Two layer, lower layer parts：Upper layer is cultivation region 21, includes specifically liq culture medium；Lower layer is detection zone 22, built-in sensing
Device.The specifically liq culture medium can be herein specifically green lactose bile salt broth, for detecting coliform.Institute
The device preparation process for stating quickly detection microorganism is as follows：
1, sensor is prepared：
（1）Prepare color indicator solution：0.1wt% bromthymol blues, 0.01wt% cresol reds are dissolved in 20% ethanol solution
In；
（2）Mix organosilicon material and color indicator solution：Weigh the cyclic annular first that hydrogen content is 0.32% successively first
Base hydrogen polysiloxanes 0.48g, methyl vinyl MQ silicon resin 29.25g and 30ppm platinum-alkynes base complex 0.27g are in plastic cup
In, then press organosilicon material：Color indicator solution=3:0.5 mass ratio weighs color indicator solution in same plastics
It in cup, is uniformly mixed, indicator is made to be dispersed in organosilicon material, and be sub-packed in bottle bottom with cover, about 1cm is thick
Degree is coagulation forming after standing 1-24 hours.
2, filling and sterilized liquid culture medium：
A. green lactose bile salt broth is poured into sensor in the bottle of coagulation forming, the body of green lactose bile salt broth
Product is the 9/10 of bottle volume；
B. after covering bottle cap, bottle is put into high-pressure sterilizing pot and is sterilized by the sterilizing requirement of nutrient broth.
C. finished product can be obtained after taking out.
Using：When using this microbial detection device, sample is subjected to increasing bacterium with suitable fluid nutrient medium first, so
After take 0.1-1mL sample enrichment liquids to be added in the device, tighten liquid in bottle cap mixing bottle and be just placed at 36 DEG C and cultivate 1-
24h.The sensor of the bottom of device turns yellow, and indicates that the sample added dilutes the presence for having coliform in even liquid.Being added should
The quantity of objective microbe contained by sample in device is more, and the time used in sensor discoloration is shorter.
Embodiment 4
As shown in Figure 1, a kind of device of quick detection microorganism of the present invention, including bottle cap 1 and body 2, it is upper that body 2, which divides,
Two layer, lower layer parts：Upper layer is cultivation region 21, includes specifically liq culture medium；Lower layer is detection zone 22, built-in sensing
Device.The specifically liq culture medium can be herein specifically RV meat soups, for detecting salmonella.The quick detection
The device preparation process of microorganism is as follows：
1, the preparation of sensor：
（1）Prepare color indicator solution：1wt% bromthymol blues, 0.1wt% xylenol blues are dissolved in 20% ethanol solution
In；
（2）Mix organosilicon material and color indicator solution：Weigh the methyl hydrogen that hydrogen content is 0.02% successively first
MQ silicones 6g, α, ω-divinyl polydimethyl silicone oil 23.4g and 30ppm platinum-alkynes base complex 0.6g is in plastic cup
In, then press organosilicon material：Color indicator solution=3:0.15 mass ratio weighs color indicator solution in same plastics
It in cup, is uniformly mixed, indicator is made to be dispersed in organosilicon material, and be sub-packed in bottle bottom with cover, about 1cm is thick
Degree is coagulation forming after standing 1-24 hours.
2, filling and sterilized liquid culture medium：
A. RV meat soups are poured into sensor in the bottle of coagulation forming, the volume of RV meat soups is the 7/10 of bottle volume；
B. after covering bottle cap, bottle is put into high-pressure sterilizing pot and is sterilized by the sterilizing requirement of nutrient broth；
C. finished product can be obtained after taking out.
Using：When using this microbial detection device, sample is subjected to increasing bacterium with suitable fluid nutrient medium first, so
After take 0.1-1mL sample enrichment liquids to be added in the device, tighten liquid in bottle cap mixing bottle and be just placed at 36 DEG C and cultivate 1-
24h.The sensor of the bottom of device turns yellow, and indicates that the sample added dilutes the presence for having salmonella in even liquid.Being added should
The quantity of objective microbe contained by sample in device is more, and the time used in sensor discoloration is shorter.
Embodiment 5
As shown in Figure 1, a kind of device of quick detection microorganism of the present invention, including bottle cap 1 and body 2, it is upper that body 2, which divides,
Two layer, lower layer parts：Upper layer is cultivation region 21, includes specifically liq culture medium；Lower layer is detection zone 22, built-in sensing
Device.The specifically liq culture medium can be herein specifically green lactose bile salt broth, for detecting coliform.Institute
The device preparation process for stating quickly detection microorganism is as follows：
1, sensor is prepared：
（1）Prepare color indicator solution：0.65wt% bromthymol blues are dissolved in 20% ethanol solution；
（2）Mix organosilicon material and color indicator solution：Weigh the linear first that hydrogen content is 0.32% successively first
Base hydrogen polysiloxanes 0.48g, α, ω-divinyl polydimethyl silicone oil 29.25g and 30ppm platinum-alkynes base complex 0.27g
In plastic cup, organosilicon material is then pressed：Color indicator solution=3:0.22 mass ratio weigh color indicator solution in
In same plastic cup, it is uniformly mixed, so that indicator is dispersed in organosilicon material, and be sub-packed in bottle bottom with cover,
About 1cm thickness is coagulation forming after standing 1-24 hours.
2, filling and sterilized liquid culture medium：
A. green lactose bile salt broth is poured into sensor in the bottle of coagulation forming, the body of green lactose bile salt broth
Product is the 4/5 of bottle volume；
B. after covering bottle cap, bottle is put into high-pressure sterilizing pot and is sterilized by the sterilizing requirement of nutrient broth；
C. finished product can be obtained after taking out.
Using：When using this microbial detection device, sample is subjected to increasing bacterium with suitable fluid nutrient medium first, so
After take 0.1-1mL sample enrichment liquids to be added in the device, tighten liquid in bottle cap mixing pipe and be just placed at 36 DEG C and cultivate 1-
24h.The sensor of the bottom of device turns yellow, and indicates that the sample added dilutes the presence for having coliform in even liquid.Being added should
The quantity of objective microbe contained by sample in device is more, and the time used in sensor discoloration is shorter.
Embodiment 6
This place sample product is ice material class cold drink, limitation requirement total plate count ﹤ 100CFU/g, therefore the microorganism inspection used
The fluid nutrient medium surveyed in device is nutrient broth.Detecting step is as follows：
(1) sample is handled：Primary sample is diluted with sterile saline or phosphate buffer, is made 1:10
The even liquid of sample；
(2) pipette in the device through quickly detecting microorganism made from step (1) treated sample 0.1mL to embodiment 1,
It is uniformly mixed after sealing；
(3) cultivate：It is 1-24 hours that the device of the quick detection microorganism inspection of step (2), which is placed on culture at 36 DEG C,；
(4) judging result：Whether visually observe sensor priming color in the device of the quick detection microorganism of step (3)
Hair
Raw to change, what priming color changed then shows there is bacterium in sample, total number of bacteria ﹥ 100CFU/ in primary sample
G, failed sample；What priming color did not change then shows do not have bacterium in sample, total number of bacteria ﹤ in primary sample
100CFU/g, sample passes.
Table 2
Used sample is all purchased from supermarket in table 2,30 DEG C of placement 12h in the case where keeping Packing Intact, present invention detection
As a result the detection duration in is the time for starting to observe and record at interval of 2~5h after culture, substantially understands that bacterium contains in primary sample
Amount is higher, and detection duration is shorter, and testing result comes out more early.In addition new light sources and photon detection technology can be used, use
Computer control system carrys out light transmittance or the index of refraction variation of complete monitoring sensor, obtains testing result in real time.
Embodiment 7
This place sample product is Yoghourt, 10 CFU/g of limitation requirement mould saccharomycete ﹤, therefore the microorganism detection used is set
In fluid nutrient medium be malt juice liquid medium.Detecting step is as follows：
(1) sample is handled：Primary sample is diluted with sterile saline or phosphate buffer, is made 1:10
The even liquid of sample；
(2) pipette in the device through quickly detecting microorganism made from step (1) treated sample 1mL to embodiment 2, it is close
It is honored as a queen uniformly mixed；
(3) cultivate：It is 1-48 hours that the device of the quick detection microorganism of step (2), which is placed on culture at 28 DEG C,；
(4) judging result：Whether visually observe sensor priming color in the device of the quick detection microorganism of step (3)
Hair
Raw to change, what priming color changed then shows there is bacterium in sample, total number of bacteria ﹥ 10CFU/g in primary sample,
Failed sample；What priming color did not change then shows do not have bacterium in sample, total number of bacteria ﹤ 10CFU/ in primary sample
G, sample passes.
Table 3
Used sample is all purchased from supermarket in table 3, is placed 4 days for 28 DEG C in the case where keeping Packing Intact.The present invention detects
As a result the detection duration in is the time for starting to observe and record at interval of 2~5h after culture, substantially understands that bacterium contains in primary sample
Amount is higher, and detection duration is shorter, and testing result comes out more early.In addition new light sources and photon detection technology can be used, use
Computer control system comes complete monitoring sensor light transmittance or index of refraction variation, obtains testing result in real time.
Embodiment 8
This place sample product is pasteurize milk, and limitation requirement is salmonella ﹤ 0/25g, therefore the microorganism inspection used
The fluid nutrient medium surveyed in setting is RV meat soups.Detecting step is as follows：
(1) sample is handled：It takes 25g Pasteur milk sample to be mixed with one step enriched medium of 225mL salmonellas and is placed in 42 DEG C of increasings
Bacterium cultivates 18-24h；
(2) the device through quickly detecting microorganism made from step (1) treated sample 0.1-1mL to embodiment 4 is pipetted
In, it is uniformly mixed after sealing；
(3) cultivate：It is 1-24 hours that the device of the quick detection microorganism of step (2), which is placed on incubation time at 36 DEG C,
(4) judging result：Whether visually observe sensor priming color in the device of the quick detection microorganism of step (3)
It changes, what priming color changed then shows, with the presence of bacterium, to need to carry out the culture solution in detection device in sample
The biochemical identification of lower step, finally to determine whether sample is exceeded；What priming color did not change then shows not having in sample
Bacterium exists, sample passes.
Table 4
Used sample is all purchased from supermarket in table 4, due to can't check salmonella substantially in bus sterilization milk on the market, looks for
Less than positive, therefore fresh addition 0.1mL salmonella in 25mL buses sterilize milk 3 and 25mL buses sterilization milk 4
Culture, as positive.Detection duration in testing result of the present invention is observed and recorded at interval of 2~5h after starting culture
Time, substantially known to bacterial content is higher in primary sample, detection duration is shorter, and testing result comes out more early.In addition may be used
Using new light sources and photon detection technology, become come complete monitoring sensor light transmittance or index of refraction using computer control system
Change, obtains testing result in real time.
Claims (7)
1. a kind of method for rapid inspecting animalcule, it is characterised in that：The target in specifically liq culture medium enrichment culture sample
During the entire process of microorganism, target is detected by organosilicon material and the sensor of color indicator solution composition using a kind of
The small molecule product that microorganism is metabolized when growing, such metabolite, which penetrates into sensor, causes the color of sensor to be sent out
Changing to whether there is objective microbe in judgement sample with this；The organosilicon material includes that hydrogen content is 0.24%
Linear methyl hydrogen polysiloxanes, α, ω-divinyl polydimethyl silicone oil and 10ppm platinum-vinyl siloxane complexs,
Weight ratio is 0.8：28.5：0.7；The content of color indicator is 0.01wt%-1wt% in the color indicator solution；Institute
The weight ratio for stating organosilicon material and color indicator solution is 3:0.2；This method includes preparing a kind of quickly detection microorganism
Device, the device include upper layer, lower layer's two parts, the specifically liq culture medium built in upper layer, the biography built in lower layer
Sensor, the sensor are in direct contact with the specifically liq culture medium.
2. a kind of method for rapid inspecting animalcule described in accordance with the claim 1, which is characterized in that the specifically liq training
The effect for supporting base is that abundant nutriment is provided for objective microbe, promotes growth breeding, while inhibiting non-targeted micro- life
The growth of object, the specifically liq culture medium using nutrient broth, pancreas peptone soybean broth, malt juice liquid medium,
Green lactose bile salt broth, AC meat soups, Bu Shi-Haas meat soup, potato dextrose broth, PA meat soups, RV meat soupsIn it is arbitrary It is a kind of.
3. a kind of method for rapid inspecting animalcule described in accordance with the claim 1, which is characterized in that the color indicator is selected from
Bromocresol green, phenolphthalein, phenol red, cresol red, reindeer moss, thymol blue, bromthymol blue, pyrocatechol purple, azolitmin, quinone quinoline are sub-
Amine, bright orange, dimethyl diaminophenazine chloride, rosolic acid, rheochrysin, 2,2 '-dihydroxy benzenes ketenes, alphanaphthol phthalein, meta cresol purple, curcumin, two
It is one or more in cresols basket.
4. a kind of method for rapid inspecting animalcule described in accordance with the claim 1, which is characterized in that the sensor is impermeable
Bright or translucent elastic solid, priming color is black, brown, dark brown or blackish green, because organosilicon material has high breathe freely
Property and selectively certain gases and small-molecule substance is only allowed to pass through, and liquid, solid and microorganism cannot all enter, therefore
The interference of sample can be significantly excluded, the only small-molecule substance in microorganism growth metabolism product can just penetrate into biography
In sensor and the color of sensor is caused to eventually become yellow, red, orange red or orange-yellow, and make sensor light transmittance or
Index of refraction increases；The microorganism small molecule metabolite that the sensor detects is more, its discoloration time used gets over
Short, the light transmittance or index of refraction of sensor are bigger.
5. a kind of method for rapid inspecting animalcule described in accordance with the claim 1, which is characterized in that the quickly detection of described one kind
The device of microorganism is the transparent resealable container of cylinder, square or cuboid.
6. a kind of method for rapid inspecting animalcule described in accordance with the claim 1, which is characterized in that including step used below：
(1) sample is handled：Sample is done into appropriate dilution with sterile diluent, fluid sample is handled without dilution；
(2) pipette in the device in right amount through step (1) treated sample to quickly detection microorganism as claimed in claim 5,
It is uniformly mixed after sealing；
(3) cultivate：The device of the quick detection microorganism of step (2) is placed at the suitable temperature of objective microbe and is cultivated, bacterium
The culture 1-24h of class, the culture 1-48h of Mycophyta；
(4) judging result：Whether the sensor priming color visually observed in the device of the quick detection microorganism of step (3) occurs
Change, what priming color changed then shows not changing then with the presence of objective microbe, priming color in sample
Show there is no objective microbe presence in sample.
7. a kind of method for rapid inspecting animalcule according to claim 6, which is characterized in that (4) the step uses automatic
The realization means of change, i.e.,:Using external light source and photon detection technology to sensor light transmittance caused by color change or folding
The variation of light rate monitors in real time, is automatically analyzed by computer control system and quickly determines objective microbe content.
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| CN108254348B (en) * | 2018-01-31 | 2020-11-17 | 国家烟草质量监督检验中心 | pH sensitive fluorescent sensor for high-throughput detection of active microorganisms and construction method |
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| CN110804644A (en) * | 2019-11-22 | 2020-02-18 | 阮雁春 | Method and device for detecting multiple microorganisms in food |
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