CN106350575A - Embossing method for bacterium sampling - Google Patents
Embossing method for bacterium sampling Download PDFInfo
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- CN106350575A CN106350575A CN201610780817.4A CN201610780817A CN106350575A CN 106350575 A CN106350575 A CN 106350575A CN 201610780817 A CN201610780817 A CN 201610780817A CN 106350575 A CN106350575 A CN 106350575A
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- culture dish
- bacterial sampling
- neutralizer
- stamped method
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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
- C12Q1/22—Testing for sterility conditions
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/10—Petri dish
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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Abstract
The invention relates to bacterium sampling method, and in particular to an embossing method for bacterium sampling. The embossing method for bacterium sampling comprises the following steps: firstly, preparing a neutralization liquid, mixing the neutralization liquid with nutrient agar, and sterilizing so as to obtain a mixed liquid; pouring the mixed liquid into a culture dish, cooling, and performing bacterium sampling by using embossing method, culturing, and calculating the clump count. The embossing method provided by the invention is convenient, simple and rapid, is capable of effectively improving the working efficiency, can be not only used for evaluating hand sterilization effects of hospitals, but also used for sampling bacteria on surfaces of articles, and has very high popularization values; when culture dish sampling is implemented by using the embossing method, a culture medium can sufficiently contact a hand or an environment surface, the culture counting process is sealed, no pollution can be caused, and analysis results can be relatively accurate.
Description
Technical field
The present invention relates to a kind of method of bacterial sampling is and in particular to a kind of stamped method for bacterial sampling.
Background technology
Body surface, medical worker bacterial population on hand and evaluation methodology are countries according to " hospital disinfection sanitary standard "
Common detection methods, require according to national standard, and all clinical line section office of all types hospital are monthly required for sampling one
Secondary, and the staff of each section office is required for carrying out sample detecting, and sampled point is many, frequency is high, sampling process is complicated, impact
Sampled result many factors.
According to the requirement of Medical personnel hand hygienic practice, after sterilization, rear examinee's the five fingers close up, and are neutralized containing corresponding with being soaked with
The cotton swab that the aseptic eluent of liquid soaks, refers to curved surface from referring to finger tip double rub 2 times, handss embrocate area in both hands
About 30cm2, rotates cotton swab simultaneously during embrocating, no special circumstances experiment needs to embrocate two gesture and facial expressions and amasss as 60cm2；By cotton
The part of swab operating of contacts person cuts off, and puts in the aseptic eluent test tube that 10ml contains corresponding nertralizer, timely censorship.
This process workload is big, is unfavorable for improving the enthusiasm that medical worker participates in, indirectly reduce hand hygiene and
Sanitary supervision.Program very complicated, it is impossible to ensure that each Bu Doushi sterile working, easily causes sampled result simultaneously
Inaccurate.
Content of the invention
According to deficiency of the prior art above, it is an object of the invention to provide a kind of stamped method for bacterial sampling,
Can be used in the evaluation of hospital's hand-wrist bones effect moreover it is possible to be applied to body surface bacterial sampling, sampling is quick, and result is more accurate.
Stamped method for bacterial sampling of the present invention, first, prepares neutralizer, then by neutralizer and nutrition fine jade
Fat mixes, sterilizing, obtains mixed liquor；Mixed liquor is poured in culture dish, after cooling, bacterial sampling is carried out using stamped method, culture,
Count clump count.
Form culture medium after neutralizer and the mixed liquor cooling of Nutrient agar.
Described neutralizer is adopted and is prepared with the following method: by soybean lecithin, Tween 80, peptone, sodium chloride, thio sulfur
Sour sodium and glycine are added to the water, and are subsequently adding ph regulator and adjust ph to 7.2-7.4, obtain neutralizer.
In described neutralizer, the percentage ratio that each component accounts for the quality of water is respectively as follows:
Soybean lecithin 0.2-0.4%, Tween 80 0.3-1%, peptone 0.8-1.2%, sodium chloride 0.5-0.85%,
Sodium thiosulfate 0.1-0.2%, glycine 0.3-0.4%.
Described ph regulator is one kind of sodium hydroxide, hydrochloric acid or citric acid.When preparing neutralizer, by Semen sojae atricolor lecithin
After fat, Tween 80, peptone, sodium chloride, sodium thiosulfate and glycine are added to the water, according to different formula, employing
Ph regulator is different, according to the ph value measuring, during more than 7.4, using citric acid or dilute hydrochloric acid as ph regulator, less than 7.2
When, using sodium hydroxide as ph regulator.
The volume ratio that described neutralizer is mixed with Nutrient agar is 1:4-5.
The temperature that described mixed liquor is poured into during culture dish is 40-45 DEG C.
The height that described mixed liquor pours in culture dish into is higher than the peak 1-2mm at culture dish back box edge.Culture dish
The peak at back box edge is the circular platform in back box.
Described sterilizing is to be placed in autoclave sterilizer to be sterilized, and sterilizes half an hour, high temperature is similarly hereinafter at 121 DEG C
When so that Nutrient agar is uniformly dissolved.
Described using the method that stamped method carries out bacterial sampling is: the media surface in culture dish is directly overlayed
Palm root extremely refers to the root 10-20 second.
Described culture is culture 24-48h in 35-37 DEG C of calorstat, and described method of counting is: clump count (cfu/
cm2Clump count/culture dish inner bottom disc area (cm on)=culture dish2).
With regard to the determination of incubation time, incubation time is 24-48h, when cultivating first, should in 24h, 30h, 36h, 42h,
48h is counted respectively, chooses the constant time point of clump count as incubation time.
Described culture dish includes lid and back box, and the periphery outer of back box is provided with circular platform, circular platform
Inner side sets lock, the interior chassis of back box sets arc convex, the top of arc convex sets grid lines.
Described grid lines is salient line.
The setting of circular platform, facilitates culture dish to open.Form culture medium, lock after neutralizer and Nutrient agar cooling
Setting can firmness between intensified education base and back box it is ensured that culture dish is when picking up, culture medium is unlikely to come off
Out.Arc convex is set on the interior chassis of back box, the top of arc convex sets grid lines, and described grid lines is salient line.Net
The setting of ruling can facilitate the counting of antibacterial, and the analysis of result can also may be used according to clump count divided by the area on chassis in back box
Directly read the clump count in each little lattice.Grid lines can facilitate the attachment of culture medium for salient line, pours training in mixed liquor
When in foster dish, culture fluid liquid level is unlikely to when being higher than circular platform to overflow, so that the height in culture dish poured into by mixed liquor
Higher than circular platform 1-2mm in culture dish back box it is ensured that culture medium can be adequately exposed to hand or environmental surfaces.
Lid is the shape of falling u, and the height at edge is higher than the height of circular platform, it is to avoid bacterial sampling finishes when covering lid
The inner surface of lid is contacted with culture medium, affects count of bacteria.
Lid adopts transparent material to prepare, and can observe the situation in culture dish at any time, can be right in the case of closing
Antibacterial is counted it is ensured that analysis result is more accurate.
Compared with prior art, the invention has the beneficial effects as follows:
1st, the method for the invention, convenient and simple quick, work efficiency can be effectively improved, can be not only used for hospital's handss and disappear
The evaluation of toxic effect fruit, moreover it is possible to be applied to body surface bacterial sampling, has very high promotional value.
When the 2nd, adopting culture dish sampling of the present invention, ensure that culture medium is adequately exposed to hand or environmental surfaces, culture
Counting process closing is carried out, pollution-free generation, and analysis result is more accurate.
Brief description
Fig. 1 is the structural representation of culture dish in the present invention；
Fig. 2 is the structural representation of the back box of culture dish in the present invention；
In figure: 1, lid；2nd, back box；3rd, circular platform；4th, grid lines；5th, latch；6th, arc convex.
Specific embodiment
Below in conjunction with the accompanying drawings embodiments of the invention are described further.
As shown in figure 1, described culture dish includes lid 1 and back box 2, the periphery outer of back box 2 is provided with circular platform
3, the inner side of circular platform 3 sets lock 5, the interior chassis of back box 2 sets arc convex 6, the top of arc convex 6 sets grid lines
4.
Described grid lines 4 is salient line.Lid adopts transparent material to prepare.
Embodiment 1
The described stamped method for hand bacterial sampling, comprises the steps:
(1) prepare neutralizer, soybean lecithin, Tween 80, peptone, sodium chloride, sodium thiosulfate and glycine are added
Enter in water, be subsequently adding sodium hydroxide solution and adjust ph to 7.3, obtain neutralizer.
In neutralizer, the percentage ratio that each component accounts for the quality of water is respectively as follows: soybean lecithin 0.3%, Tween 80 0.3%,
Peptone 1%, sodium chloride 0.5%, sodium thiosulfate 0.1%, glycine 0.3%.
(2), after neutralizer being mixed homogeneously with the Nutrient agar of dissolving, it is placed in autoclave sterilizer and is sterilized, obtain mixed
Close liquid.
(3) mixed liquor is cooled to after 42 DEG C, pours in culture dish, cooling is stand-by, and the mixed liquor after cooling becomes culture medium.
(4) media surface on culture dish is directly overlayed after palm root extremely refers to root 15 seconds, cultivate 24h, according to
Grid on culture dish counts.
The amount that mixed liquor pours in culture dish into is higher than circular platform 1mm.
Neutralizer is 1:4 with the volume ratio of the Nutrient agar of dissolving.
Embodiment 2
The described stamped method for body surface bacterial sampling, comprises the steps:
(1) prepare neutralizer, soybean lecithin, Tween 80, peptone, sodium chloride, sodium thiosulfate and glycine are added
Enter in water, be subsequently adding dilute hydrochloric acid solution and adjust ph to 7.2, obtain neutralizer.
In neutralizer, the percentage ratio that each component accounts for the quality of water is respectively as follows: soybean lecithin 0.2%, Tween 80 0.4%,
Peptone 0.8%, sodium chloride 0.6%, sodium thiosulfate 0.15%, glycine 0.4%.
(2), after neutralizer being mixed homogeneously with the Nutrient agar of dissolving, it is placed in autoclave sterilizer and is sterilized, obtain mixed
Close liquid.
(3) mixed liquor is cooled to after 45 DEG C, pours in culture dish, cooling is stand-by, and the mixed liquor after cooling becomes culture medium.
(4) when detecting, the surface of culture dish was overlayed in tested object upper 20 second, cultivate 48h, according to the net on culture dish
Lattice count.
The amount that mixed liquor pours in culture dish into is higher than circular platform 2mm.
Neutralizer is 1:5 with the volume ratio of the Nutrient agar of dissolving.
Claims (10)
1. a kind of stamped method for bacterial sampling it is characterised in that: first, prepare neutralizer, then by neutralizer and nutrition
Agar mixes, sterilizing, obtains mixed liquor；Mixed liquor is poured in culture dish, after cooling, bacterial sampling is carried out using stamped method, training
Support, count clump count.
2. the stamped method for bacterial sampling according to claim 1 it is characterised in that: described neutralizer is using as follows
Method is prepared: soybean lecithin, Tween 80, peptone, sodium chloride, sodium thiosulfate and glycine is added to the water, then
Add ph regulator to adjust ph to 7.2-7.4, obtain neutralizer.
3. the stamped method for bacterial sampling according to claim 2 it is characterised in that: in described neutralizer, each group
The percentage ratio dividing the quality accounting for water is respectively as follows:
Soybean lecithin 0.2-0.4%, Tween 80 0.3-1%, peptone 0.8-1.2%, sodium chloride 0.5-0.85%, thio
Sodium sulfate 0.1-0.2%, glycine 0.3-0.4%.
4. the stamped method for bacterial sampling according to claim 2 it is characterised in that: described ph regulator be hydrogen-oxygen
Change one kind of sodium, hydrochloric acid or citric acid.
5. the stamped method for bacterial sampling according to claim 1 it is characterised in that: described neutralizer and nutrition fine jade
The volume ratio of fat mixing is 1:4-5.
6. the stamped method for bacterial sampling according to claim 1 it is characterised in that: culture poured into by described mixed liquor
During dish, temperature is 40-45 DEG C.
7. the stamped method for bacterial sampling according to claim 1 it is characterised in that: culture poured into by described mixed liquor
Height in dish is higher than the peak 1-2mm at culture dish back box edge.
8. the stamped method for bacterial sampling according to claim 1 it is characterised in that: described is carried out using stamped method
The method of bacterial sampling is: the media surface in culture dish is directly overlayed and extremely refers to the root 10-20 second in palm root.
9. the stamped method for bacterial sampling according to claim 1 it is characterised in that: described culture is in 35-37
24-48h is cultivated, described method of counting is: clump count (cfu/cm in DEG C calorstat2Clump count/culture dish on)=culture dish
Inner bottom disc area (cm2).
10. the stamped method for bacterial sampling according to claim 1 it is characterised in that: described culture dish includes box
Lid (1) and back box (2), the periphery outer of back box (2) is provided with circular platform (3), and the inner side of circular platform (3) sets lock
(5), the interior chassis of back box (2) sets arc convex (6), the top of arc convex (6) sets grid lines (4), described grid lines
(4) it is salient line.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610780817.4A CN106350575A (en) | 2016-08-31 | 2016-08-31 | Embossing method for bacterium sampling |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610780817.4A CN106350575A (en) | 2016-08-31 | 2016-08-31 | Embossing method for bacterium sampling |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106350575A true CN106350575A (en) | 2017-01-25 |
Family
ID=57856265
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610780817.4A Pending CN106350575A (en) | 2016-08-31 | 2016-08-31 | Embossing method for bacterium sampling |
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| CN (1) | CN106350575A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112300906A (en) * | 2020-10-21 | 2021-02-02 | 上海源培生物科技股份有限公司 | Finger sampling culture dish |
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| CN2606111Y (en) * | 2003-04-01 | 2004-03-10 | 济南市疾病预防控制中心 | Culture plate for quick bacteria sample |
| CN101698864A (en) * | 2007-12-18 | 2010-04-28 | 曲奕 | Air sampling neutralization culture medium in public sanitary place |
| CN102928553A (en) * | 2011-08-09 | 2013-02-13 | 崔瑛 | Disinfection effect experiment neutralizer set |
| CN202188973U (en) * | 2011-09-05 | 2012-04-11 | 中国人民解放军济南军区联勤部疾病预防控制中心 | Injector type agar stamping sampler |
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| CN104364388A (en) * | 2012-04-16 | 2015-02-18 | 快速微型生物系统公司 | Cell culturing device |
| CN104312915A (en) * | 2014-10-27 | 2015-01-28 | 济南百博生物技术股份有限公司 | Compound neutralizing and sterilizing monitoring device and method for monitoring by using same |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112300906A (en) * | 2020-10-21 | 2021-02-02 | 上海源培生物科技股份有限公司 | Finger sampling culture dish |
| CN112300906B (en) * | 2020-10-21 | 2023-01-03 | 上海源培生物科技股份有限公司 | Finger sampling culture dish |
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