CN107151688A - In a kind of public sanitary place air sampling and culture medium - Google Patents
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- CN107151688A CN107151688A CN201710365784.1A CN201710365784A CN107151688A CN 107151688 A CN107151688 A CN 107151688A CN 201710365784 A CN201710365784 A CN 201710365784A CN 107151688 A CN107151688 A CN 107151688A
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
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Abstract
The invention discloses in a kind of public sanitary place air sampling of culture medium preparing technical field and culture medium, in this kind of public sanitary place air sampling and culture medium preparation method step it is as follows：S1：Chosen material；S2：Filtering, dissolving；S3：Adjust pH；S4：Filtering clarification；S5：Packing；S6：Sterilizing；S7：Calibrating and preservation, lecithin in nertralizer can neutralize quaternary ammonium compound, Tween 80 can neutralize phenolic compound, sodium thiosulfate can neutralize the preservative with oxidisability such as iodine and chlorine, sodium thioglycolate can be neutralized containing mercurial antiseptic, there is solid neutralization to residual public sanitary place air sanitizer, it can be chemically reacted with residual public sanitary place air sanitizer, the effect of its growth is not killed or suppressed in generation to experimental microbial, there is no destruction to culture composition, do not have influential neutralized reaction product on physical behavior, there is solid neutralization to corresponding residual public sanitary place air sanitizer.
Description
Technical field
The present invention relates to culture medium preparing technical field, in specially a kind of public sanitary place air sampling and cultivate
Base.
Background technology
The microorganism of public sanitary place air is influenceed by the chemical composition of disinfectant, and public place air is solved first
A variety of chemical compositions of middle disinfective action are key core parts, and need strictly to grasp its action time.This skill
Art be according to air sanitizer contacts certain time with microorganism in public sanitary place after, the microorganism in air becomes
Change evaluate air sanitizer effect so that compare market sale various air sanitizers (including indoor air disinfectant) and
Develop the exploitation and production of more effectively air sanitizer.So far, the side of residual disinfectant chemical composition in air is removed
Method is a lot, such as from CNKI 7 phases in 2016《Shrink》, the remaining sterilization delivered in August, 2016
The sweep-out method of agent, for the composition of acid, alkali present in disinfectant, i.e., is come from methods such as absorption, dilution, centrifugation, filterings
Remove chemical residue composition.But there is limitation in these methods, if the shortcoming of dilution process be the bacterium number survived in sample
Through less, the bacterium in sample can be greatly lowered after diluting again, cause not examine in sensing chamber, cause false negative
Result occur, filtration method using process it is relatively complicated, the technical requirements to operating personnel are higher, and these methods all compare
Simply, limitation is than larger, in current method using it is more be chemical neutralisation, present nertralizer species has a lot, but pin
It to different disinfectants and bacterium, there is no preferable nertralizer to be handled so far, be free of so how to take in air sample
Residual disinfectant a variety of chemical substances are that this technology will solve core, just Detection of Air Quality method can be made more accurate
Really, reliably.Therefore, we neutralize culture medium technology from novel air sampling, the air sanitizer that residual is removed comprehensively is various
Chemical composition, the effect height of the air sanitizer to being carried out disinfection in public air is investigated to cultivate bacterium present in air
It is low, so that evaluating the air quality in public hygienic environment.
The content of the invention
It is an object of the invention to provide in a kind of public sanitary place air sampling and culture medium, to solve above-mentioned background
The problem of being proposed in technology.
To achieve the above object, the present invention provides following technical scheme：In a kind of public sanitary place air sampling and train
Base is supported, the preparation method step in this kind of public sanitary place air sampling with culture medium is as follows：
S1：Chosen material：Choose lecithin 20-30g, Tween-80 20-60g, histidine 2-10g, sodium thiosulfate 2-
10g, sodium thioglycolate 1-2g, glucose 30-40g, tryptone 20-30g, agar 40-50g, sodium chloride 17g；
S2：Filtering, dissolving：The material that step S1 is chosen is passed into active carbon filter successively filtered before air
Processing, then the addition of lecithin, Tween-80, histidine, sodium thiosulfate and sodium thioglycolate is heated stir in heater
Mix, mixing time 25-35 minutes, temperature setting forms solution one, then by glucose, tryptone, fine jade at 80 DEG C after dissolving
Fat and sodium chloride are inserted in heater, stirring mixing 10-20 minutes, solution two are formed after dissolving, then by solution one
The mixed solution of dissolving is added to solution two, and stirring is mixed 10-16 minutes, and it is finally mixed by what is obtained to stand cooling 10-20 minutes
The water for closing 80-120 milliliters of addition in material is sufficiently stirred for 10-20 minutes, is all uniformly dissolved to each composition with water, is formed after dissolving
Culture medium；
S3：Adjust pH：Three, the test tube with bore with standard pipe is taken, in each training for adding pH to be determined of first and third pipe
Base 5ml is supported, and addition 0.2g/L phenol red 0.25ml is managed as measure in the first pipe, is fully mixed, in the addition steaming of the second pipe
Distilled water 5ml, standard pipe is pH standard colorimetric tubes, and then the accurate slow solution that will correct is corrected to pipe is determined, until face
Untill color is identical with standard pipe after being determined by pH test paper, color is in lavender, and its pH value is adjusted to 7, is all needed after often Jia one and dripping
Fully to mix, Jia the second again after colorimetric drips the amount that accurate recording is added；
S4：Filtering clarification：Culture medium in step S2 is imported into container, 10 are melted with high pressure 100-110kPa steam
After minute, 12 hours overnight are stood in pressure cooker, next day pours out culture medium, is cut bottom sediment with knife, then melt
Receive clearly culture medium；
S5：Packing：The clearly culture medium heating and melting into step S4, temperature is 120-130 DEG C, and the time is 1 hour, so
Quiescent culture base 30 minutes afterwards so that temperature is cooled to 50 DEG C or so in culture medium, is poured into sterile formality in sterilizing plates, plate
The culture medium poured into about 13-15mol/L, jog plate bottom makes culture medium be laid in plate bottom, to be solidified rear standby；
S6：Sterilizing：High steam is sterilized after the culture medium that will be solidified in step S5, and temperature is 115 DEG C, and the time is
15-30min；
S7：Calibrating and preservation：Culture medium must can be used after being made through calibrating, put culture medium in 37 DEG C of incubators during calibrating
Culture 24 hours, the culture medium made is placed on standby in 4-6 DEG C of refrigerator.
It is preferred that, the heater in the step S2 is electric furnace.
It is preferred that, the correction liquid in the step S3 is peracid or aqueous slkali, and peracid or aqueous slkali use excessively excessively
0.1mol/L hydrochloric acid solutions or 0.1mol/L sodium hydroxides.
It is preferred that, the container in the step S4 is aluminum pot or wide-mouth enamelled vessel.
It is preferred that, the internal diameter of the plate in the step S5 is nine centimetres.
Compared with prior art, the beneficial effects of the invention are as follows：In this kind of public sanitary place air sampling and culture medium
Lecithin in nertralizer can neutralize quaternary ammonium compound, and Tween-80 can neutralize phenolic compound, and sodium thiosulfate, which can be neutralized, to be had
The preservative of oxidisability such as iodine and chlorine, sodium thioglycolate can be neutralized containing mercurial antiseptic, and histidine can neutralize aldehydes preservative, above-mentioned
Material has solid neutralization after mixing in proportion to residual public sanitary place air sanitizer, can be public with residual
Altogether sanitary units air sanitizer chemically react, generation experimental microbial is not killed or suppress its grow effect,
There is no destruction to culture composition, do not have influential neutralized reaction product on physical behavior, it is empty to corresponding residual public sanitary place
Gas disinfectant has solid neutralization.
Brief description of the drawings
Fig. 1 is workflow diagram of the present invention；
Fig. 2 is the bacterial concentration schematic diagram after the general medium treatment of the present invention；
Fig. 3 is the bacterial concentration schematic diagram after medium treatment of the present invention；
Fig. 4 is bacterium of the present invention in medium treatment cross-reference figure.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation is described, it is clear that described embodiment is only a part of embodiment of the invention, rather than whole embodiments.It is based on
Embodiment in the present invention, it is every other that those of ordinary skill in the art are obtained under the premise of creative work is not made
Embodiment, belongs to the scope of protection of the invention.
Embodiment one
In a kind of public sanitary place air sampling and culture medium, in this kind of public sanitary place air sampling and culture medium
Preparation method step it is as follows：
S1：Chosen material：Choose lecithin 20g, Tween-80 20g, histidine 2g, sodium thiosulfate 2g, sodium thioglycolate
1g, glucose 30g, tryptone 20g, agar 40g, sodium chloride 17g；
S2：Filtering, dissolving：The material that step S1 is chosen is passed into active carbon filter successively filtered before air
Processing, then the addition of lecithin, Tween-80, histidine, sodium thiosulfate and sodium thioglycolate is heated stir in heater
Mix, mixing time 25 minutes, temperature setting forms solution one, then by glucose, tryptone, agar at 80 DEG C after dissolving
Inserted with sodium chloride in heater, stirring mixing 10 minutes forms solution two after dissolving, then by dissolving in solution one
Mixed solution is added to solution two, and stirring mixing 10 minutes, standing cooling will finally add 80 in 10 minutes in obtained compound
The water of milliliter is sufficiently stirred for 10 minutes, is all uniformly dissolved to each composition with water, culture medium is formed after dissolving；
S3：Adjust pH：Three, the test tube with bore with standard pipe is taken, in each training for adding pH to be determined of first and third pipe
Base 5ml is supported, and addition 0.2g/L phenol red 0.25ml is managed as measure in the first pipe, is fully mixed, in the addition steaming of the second pipe
Distilled water 5ml, standard pipe is pH standard colorimetric tubes, and then the accurate slow solution that will correct is corrected to pipe is determined, until face
Untill color is identical with standard pipe after being determined by pH test paper, color is in lavender, and its pH value is adjusted to 7, is all needed after often Jia one and dripping
Fully to mix, Jia the second again after colorimetric drips the amount that accurate recording is added；
S4：Filtering clarification：Culture medium in step S2 is imported into container, melted 10 minutes with high pressure 100kPa steam
Afterwards, 12 hours overnight are stood in pressure cooker, next day pours out culture medium, is cut bottom sediment with knife, then melts and can receive clear
Clear culture medium；
S5：Packing：The clearly culture medium heating and melting into step S4, temperature is 120 DEG C, and the time is 1 hour, Ran Houjing
Put culture medium 30 minutes so that temperature is cooled to 50 DEG C or so in culture medium, poured into sterile formality in sterilizing plates, plate is poured into
Culture medium about 13mol/L, jog plate bottom makes culture medium be laid in plate bottom, to be solidified rear standby；
S6：Sterilizing：High steam is sterilized after the culture medium that will be solidified in step S5, and temperature is 115 DEG C, and the time is
15min；
S7：Calibrating and preservation：Culture medium must can be used after being made through calibrating, put culture medium in 37 DEG C of incubators during calibrating
Culture 24 hours, the culture medium made is placed on standby in 4 DEG C of refrigerators.
Embodiment two
In a kind of public sanitary place air sampling and culture medium, in this kind of public sanitary place air sampling and culture medium
Preparation method step it is as follows：
S1：Chosen material：Choose lecithin 25g, Tween-80 40g, histidine 6g, sodium thiosulfate 6g, sodium thioglycolate
1.5g, glucose 35g, tryptone 25g, agar 45g, sodium chloride 17g；
S2：Filtering, dissolving：The material that step S1 is chosen is passed into active carbon filter successively filtered before air
Processing, then the addition of lecithin, Tween-80, histidine, sodium thiosulfate and sodium thioglycolate is heated stir in heater
Mix, mixing time 30 minutes, temperature setting forms solution one, then by glucose, tryptone, agar at 80 DEG C after dissolving
Inserted with sodium chloride in heater, stirring mixing 15 minutes forms solution two after dissolving, then by dissolving in solution one
Mixed solution is added to solution two, and stirring mixing 13 minutes, standing cooling will finally add 100 in 15 minutes in obtained compound
The water of milliliter is sufficiently stirred for 15 minutes, is all uniformly dissolved to each composition with water, culture medium is formed after dissolving；
S3：Adjust pH：Three, the test tube with bore with standard pipe is taken, in each training for adding pH to be determined of first and third pipe
Base 5ml is supported, and addition 0.2g/L phenol red 0.25ml is managed as measure in the first pipe, is fully mixed, in the addition steaming of the second pipe
Distilled water 5ml, standard pipe is pH standard colorimetric tubes, and then the accurate slow solution that will correct is corrected to pipe is determined, until face
Untill color is identical with standard pipe after being determined by pH test paper, color is in lavender, and its pH value is adjusted to 7, is all needed after often Jia one and dripping
Fully to mix, Jia the second again after colorimetric drips the amount that accurate recording is added；
S4：Filtering clarification：Culture medium in step S2 is imported into container, melted 10 minutes with high pressure 105kPa steam
Afterwards, 12 hours overnight are stood in pressure cooker, next day pours out culture medium, is cut bottom sediment with knife, then melts and can receive clear
Clear culture medium；
S5：Packing：The clearly culture medium heating and melting into step S4, temperature is 125 DEG C, and the time is 1 hour, Ran Houjing
Put culture medium 30 minutes so that temperature is cooled to 50 DEG C or so in culture medium, poured into sterile formality in sterilizing plates, plate is poured into
Culture medium about 14mol/L, jog plate bottom makes culture medium be laid in plate bottom, to be solidified rear standby；
S6：Sterilizing：High steam is sterilized after the culture medium that will be solidified in step S5, and temperature is 115 DEG C, and the time is
22.5min；
S7：Calibrating and preservation：Culture medium must can be used after being made through calibrating, put culture medium in 37 DEG C of incubators during calibrating
Culture 24 hours, the culture medium made is placed on standby in 5 DEG C of refrigerators.
Embodiment three
In a kind of public sanitary place air sampling and culture medium, in this kind of public sanitary place air sampling and culture medium
Preparation method step it is as follows：
S1：Chosen material：Choose lecithin 30g, Tween-80 60g, histidine 10g, sodium thiosulfate 10g, TGA
Sodium 2g, glucose 40g, tryptone 30g, agar 40-50g, sodium chloride 17g；
S2：Filtering, dissolving：The material that step S1 is chosen is passed into active carbon filter successively filtered before air
Processing, then the addition of lecithin, Tween-80, histidine, sodium thiosulfate and sodium thioglycolate is heated stir in heater
Mix, mixing time 35 minutes, temperature setting forms solution one, then by glucose, tryptone, agar at 80 DEG C after dissolving
Inserted with sodium chloride in heater, stirring mixing 20 minutes forms solution two after dissolving, then by dissolving in solution one
Mixed solution is added to solution two, and stirring mixing 16 minutes, standing cooling will finally add 120 in 20 minutes in obtained compound
The water of milliliter is sufficiently stirred for 20 minutes, is all uniformly dissolved to each composition with water, culture medium is formed after dissolving；
S3：Adjust pH：Three, the test tube with bore with standard pipe is taken, in each training for adding pH to be determined of first and third pipe
Base 5ml is supported, and addition 0.2g/L phenol red 0.25ml is managed as measure in the first pipe, is fully mixed, in the addition steaming of the second pipe
Distilled water 5ml, standard pipe is pH standard colorimetric tubes, and then the accurate slow solution that will correct is corrected to pipe is determined, until face
Untill color is identical with standard pipe after being determined by pH test paper, color is in lavender, and its pH value is adjusted to 7, is all needed after often Jia one and dripping
Fully to mix, Jia the second again after colorimetric drips the amount that accurate recording is added；
S4：Filtering clarification：Culture medium in step S2 is imported into container, melted 10 minutes with high pressure 110kPa steam
Afterwards, 12 hours overnight are stood in pressure cooker, next day pours out culture medium, is cut bottom sediment with knife, then melts and can receive clear
Clear culture medium；
S5：Packing：The clearly culture medium heating and melting into step S4, temperature is 130 DEG C, and the time is 1 hour, Ran Houjing
Put culture medium 30 minutes so that temperature is cooled to 50 DEG C or so in culture medium, poured into sterile formality in sterilizing plates, plate is poured into
Culture medium about 15mol/L, jog plate bottom makes culture medium be laid in plate bottom, to be solidified rear standby；
S6：Sterilizing：High steam is sterilized after the culture medium that will be solidified in step S5, and temperature is 115 DEG C, and the time is
30min；
S7：Calibrating and preservation：Culture medium must can be used after being made through calibrating, put culture medium in 37 DEG C of incubators during calibrating
Culture 24 hours, the culture medium made is placed on standby in 6 DEG C of refrigerators.
General culture medium suppresses the growth of bacterium, causes bacterial concentration not in disinfectant and insufficient as shown in Figure 2
It can increase, be drawn from the figure 3, it may be seen that being summarized according to the experimental result of embodiment one, two, three：Optimum implementation is embodiment
Two, in embodiment two to neutralize culture medium bright with respect to the rate of recovery of embodiment one and the neutralization culture medium of embodiment two its microorganism
It is aobvious higher, and embodiment one and the speed of three kind of bacterial growth have still suppressed, growth rate is slower, so the nertralizer can
Interference of the air sanitizer to testing result is eliminated, preferably the microbe quantity in residual public sanitary place air sanitizer
Amount, improves the accuracy of the microorganism detection in residual public sanitary place air sanitizer..
Although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of changes, modification can be carried out to these embodiments, replace without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (5)
1. in a kind of public sanitary place air sampling and culture medium, it is characterised in that this kind of public sanitary place air sampling
The preparation method step for neutralizing culture medium is as follows：
S1：Chosen material：Choose lecithin 20-30g, Tween-80 20-60g, histidine 2-10g, sodium thiosulfate 2-10g, mercapto
Guanidine-acetic acid sodium 1-2g, glucose 30-40g, tryptone 20-30g, agar 40-50g, sodium chloride 17g；
S2：Filtering, dissolving：The material that step S1 is chosen is passed into active carbon filter successively to be carried out before air at filtering
Reason, then the addition of lecithin, Tween-80, histidine, sodium thiosulfate and sodium thioglycolate is heated stir in heater
Mix, mixing time 25-35 minutes, temperature setting forms solution one, then by glucose, tryptone, fine jade at 80 DEG C after dissolving
Fat and sodium chloride are inserted in heater, stirring mixing 10-20 minutes, solution two are formed after dissolving, then by solution one
The mixed solution of dissolving is added to solution two, and stirring is mixed 10-16 minutes, and it is finally mixed by what is obtained to stand cooling 10-20 minutes
The water for closing 80-120 milliliters of addition in material is sufficiently stirred for 10-20 minutes, is all uniformly dissolved to each composition with water, is formed after dissolving
Culture medium；
S3：Adjust pH：Three, the test tube with bore with standard pipe is taken, in each culture medium for adding pH to be determined of first and third pipe
5ml, and addition 0.2g/L phenol red 0.25ml is managed as measure in the first pipe, is fully mixed, in the second pipe addition distilled water
5ml, standard pipe is pH standard colorimetric tubes, and then the accurate slow solution that will correct is corrected to pipe is determined, until color is led to
Cross it is identical with standard pipe after pH test paper is determined untill, color is in lavender, and its pH value is adjusted to 7, is required for filling after often Jia one and dripping
Divide and mix, Jia the second again after colorimetric drips the amount that accurate recording is added；
S4：Filtering clarification：Culture medium in step S2 is imported into container, melted 10 minutes with high pressure 100-110kPa steam
Afterwards, 12 hours overnight are stood in pressure cooker, next day pours out culture medium, is cut bottom sediment with knife, then melts and can receive clear
Clear culture medium；
S5：Packing：The clearly culture medium heating and melting into step S4, temperature is 120-130 DEG C, and the time is 1 hour, Ran Houjing
Put culture medium 30 minutes so that temperature is cooled to 50 DEG C or so in culture medium, poured into sterile formality in sterilizing plates, plate is poured into
Culture medium about 13-15mol/L, jog plate bottom makes culture medium be laid in plate bottom, to be solidified rear standby；
S6：Sterilizing：High steam is sterilized after the culture medium that will be solidified in step S5, and temperature is 115 DEG C, and the time is 15-
30min；
S7：Calibrating and preservation：Culture medium must can be used after being made through calibrating, put in 37 DEG C of incubators culture medium during calibrating and cultivated
24 hours, the culture medium made was placed on standby in 4-6 DEG C of refrigerator.
2. in a kind of public sanitary place air sampling according to claim 1 and culture medium, it is characterised in that：The step
Heater in rapid S2 is electric furnace.
3. in a kind of public sanitary place air sampling according to claim 1 and culture medium, it is characterised in that：The step
In rapid S3 correction liquid is peracid or crosses aqueous slkali, and peracid or cross aqueous slkali using 0.1mol/L hydrochloric acid solutions or
0.1mol/L sodium hydroxides.
4. in a kind of public sanitary place air sampling according to claim 1 and culture medium, it is characterised in that：The step
Container in rapid S4 is aluminum pot or wide-mouth enamelled vessel.
5. in a kind of public sanitary place air sampling according to claim 1 and culture medium, it is characterised in that：The step
The internal diameter of plate in rapid S5 is nine centimetres.
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| CN201710365784.1A CN107151688A (en) | 2017-05-23 | 2017-05-23 | In a kind of public sanitary place air sampling and culture medium |
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