CN106755280A - Hot air sterilization biological indicator and preparation method thereof - Google Patents
Hot air sterilization biological indicator and preparation method thereof Download PDFInfo
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- CN106755280A CN106755280A CN201611127865.XA CN201611127865A CN106755280A CN 106755280 A CN106755280 A CN 106755280A CN 201611127865 A CN201611127865 A CN 201611127865A CN 106755280 A CN106755280 A CN 106755280A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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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
- C12Q1/22—Testing for sterility conditions
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention belongs to biological monitoring technical field in sterilization, and in particular to a kind of hot air sterilization biological indicator and preparation method thereof.Hot air sterilization biological indicator is constituted by sealing the ampoule bottle by bacterium piece and the plastic tube equipped with recovery media, and the two is individually present.The monitoring of hot air sterilizer sterilization effect is carried out using hot air sterilization biological indicator of the invention, simple and convenient, safe operation, visual result is accurate.
Description
Technical field
The invention belongs to biological monitoring technical field in sterilization, and in particular to a kind of hot air sterilization biological indicator and
Its preparation method.
Background technology
With the development of medical technology, new medicine equipment and medical material are continuously emerged, and people are recognized health
Knowledge level is improved constantly.In hospital's routine work, various apparatuses are required to sterilization, can after sterilizing effect monitoring is qualified
Use.The high temperature heated dry air that hot air sterilization technology is produced by the use of electrical heating is aoxidized, egg as sterilization factors to microorganism
White matter is denatured and electrolyte concentration, makes microorganism dead.The clear stipulaties in new hospital disinfection supply center management regulation,
Dry heat sterilization devices should carry out weekly a biological detection to its sterilization effect.In the checking of sterilizing program, although can pass through
Sterilization effect is assessed in the monitoring of sterilization process some parameters, but xeothermic biological indicator is then that to evaluate a sterilizing program effective
Property most intuitively index.
The content of the invention
It is easy to use it is an object of the invention to provide a kind of hot air sterilization biological indicator, as a result it is easy to observation, to xeothermic
The sterilization ability of sterilizer is preferably monitored；Invention also provides the preparation method of hot air sterilization biological indicator,
It is scientific and reasonable, simple and easy to apply.
Hot air sterilization biological indicator of the present invention is the ampoule bottle that has bacterium piece by envelope and equipped with recovery media
Plastic tube is constituted, and the two is individually present.
Described bacterium piece is made up of indicator microoraganism and bacterial carrier.
Described indicator microoraganism is bacillus subtilis black variety gemma (ATCC9372).
Described bacterial carrier is glass fiber filter paper GF/F grades.
Described ampoule bottle is Bluepoint curved neck easy breaking ampule bottle.
It is completely enclosed that ampoule bottle need to melt envelope through overheat, it is ensured that gnotobasis is in when bacterium piece is preserved, while having in sterilizing
The effect sterilizing factor can be contacted with bacterium piece.
The composition of described recovery media is as follows, and with quality volume percentage, in terms of g, volume is in terms of mL for quality：
The preparation method of described recovery media is that tryptone, soy peptone and sodium chloride are added into distilled water
In, it is well mixed to be completely dissolved, pH7.2 is adjusted, after being sterilized through 121 DEG C, obtain final product.
Described plastic tube is PET cryovials.
The preparation method of hot air sterilization biological indicator of the present invention, step is as follows：
(1) indicator microoraganism culture, spore solution is collected；
(2) dye spore solution is dripped on bacterial carrier, dried for standby obtains bacterium piece；
(3) bacterium piece is put in ampoule interior, carries out heat and melt envelope；
(4) recovery media is weighed and prepared；
(5) after recovery media sterilizing, operated in Biohazard Safety Equipment, be sub-packed in plastic tube, screw bottle cap；
(6) finished product assembling, external packing production.
Bacterial content is 1 × 10 in bacterium piece described in step (2)6Cfu/ pieces.
Technical parameter is shown in Table 1 in the preparation method of hot air sterilization biological indicator of the invention.
Technical parameter in the preparation method of the hot air sterilization biological indicator of table 1
| Performance | It is required that |
| Bacterial content | Bacterial content is 1 × 106Cfu/ pieces |
| Time-to-live | Under the conditions of 160 DEG C ± 4 DEG C of temperature, the time-to-live >=4.9min |
| The killing time | Under the conditions of 160 DEG C ± 4 DEG C of temperature, time≤19min is killed |
| D values | Under the conditions of 160 DEG C ± 4 DEG C of temperature, D values are 1.6~1.9min |
The present invention compared with prior art, has the advantages that：
(1) monitoring of hot air sterilizer sterilization effect is carried out using hot air sterilization biological indicator of the invention, simple side
Just, safe operation, visual result is accurate.
(2) gemma on hot air sterilization biological indicator bacterium piece of the invention is in drying regime, and gemma will not grow, and resists
Power stabilization, improves product quality.
(3) hot air sterilization biological indicator of the invention is used, the generation of secondary pollution can be avoided.
Brief description of the drawings
Fig. 1 is structural representation of the invention；
In figure：1st, the bacterium piece of drop dye bacillus subtilis black variety gemma；2nd, ampoule bottle；3rd, the plastics equipped with recovery media
Pipe.
Specific embodiment
The present invention is described further with reference to embodiments.
Hot air sterilization biological indicator in following examples is by sealing the ampoule bottle of bacterium piece and equipped with recovery media
Plastic tube two parts are constituted.Indicator microoraganism is bacillus subtilis black variety gemma, and bacterium piece material is glass fiber filter paper GF/F,
Ampoule bottle is 1mL Bluepoints curved neck easy breaking ampule bottle, and plastic tube is 1.5mL PET cryovials.
Embodiment 1
(1) bacillus subtilis black variety gemma culture
1. nutrient broth medium 50mL is prepared using 250ml conical flasks, used after 121 DEG C of sterilizings, choose hay bacillus
Var. niger spores 20ul is activated, soak time 5h.
2. medicine is weighed, bacillus subtilis black variety gemma growth medium is prepared, 250mL is dispensed in each gemma bottle,
Room temperature is used after being cooled to solid-state after 121 DEG C of sterilizings.
3. operated in Biohazard Safety Equipment, the bacillus subtilis black variety gemma after activation is mixed, in each gemma bottle
Inoculation 10mL, mixes and swings uniform, after inoculation liquid is fully contacted growth media surface, pours out inoculation liquid.
4. gemma bottle is put in 37 DEG C of bacteriological incubators and cultivates 48h.
5. culture receives bacterium after terminating, and 15mL sterilized waters are added in each gemma bottle, and gemma growth medium is scraped using L rods
The gemma on surface, is poured into aseptic conical flask, is repeated 3 times.
6. the spore solution being collected into carries out 3000 and leaves heart 25min, is repeated 4 times, and centrifugation is cleaned, and uses aseptic water dissolves
The gemma precipitation for finally giving.
7. bacterium amount calculating is carried out to final spore solution with dilution method, be put in 4 DEG C of Refrigerator stores.
(2) bacillus subtilis black variety gemma bacterium piece makes
1. bacterium piece is cut, and size is 3.5mm × 20mm, is used after low temperature sterilization.
2. spore solution is diluted according to first step bacterium amount count results, concentration is 1.0 × 108Cfu/mL, every bacterium piece drop
Dye 10ul, spontaneously dries overnight or more than 12h.
3. dried bacterium piece is put in Bluepoint curved neck easy breaking ampule bottle, is carried out warm high and is melted envelope.
(3) recovery media is prepared
Quality volume (g/mL) percentage of hot air sterilization biological indicator recovery media is：
Tryptone 1.5%
Soy peptone 0.5%
Sodium chloride 2%
Distilled water is prepared, well mixed to be completely dissolved, and adjusts pH7.2, is used after being sterilized through 121 DEG C.
Recovery media after sterilizing is sub-packed in aseptic plastic pipe, often addition 1.4mL in pipe, screws bottle cap, prevented
Only liquid spills, it is to avoid living contaminants.
(4) finished product assembling, external packing production.
Embodiment 2
(1) bacillus subtilis black variety gemma culture
1. nutrient broth medium 50mL is prepared using 250ml conical flasks, used after 121 DEG C of sterilizings, choose hay bacillus
Var. niger spores 20ul is activated, soak time 5h.
2. medicine is weighed, bacillus subtilis black variety gemma growth medium is prepared, 250mL is dispensed in each gemma bottle,
Room temperature is used after being cooled to solid-state after 121 DEG C of sterilizings.
3. operated in Biohazard Safety Equipment, the bacillus subtilis black variety gemma after activation is mixed, in each gemma bottle
Inoculation 10mL, mixes and swings uniform, after inoculation liquid is fully contacted growth media surface, pours out inoculation liquid.
4. gemma bottle is put in 37 DEG C of bacteriological incubators and cultivates 48h.
5. culture receives bacterium after terminating, and 15mL sterilized waters are added in each gemma bottle, and gemma growth medium is scraped using L rods
The gemma on surface, is poured into aseptic conical flask, is repeated 3 times.
6. the spore solution being collected into carries out 3000 and leaves heart 25min, is repeated 4 times, and centrifugation is cleaned, and uses aseptic water dissolves
The gemma precipitation for finally giving.
7. bacterium amount calculating is carried out to final spore solution with dilution method, be put in 4 DEG C of Refrigerator stores.
(2) bacillus subtilis black variety gemma bacterium piece makes
1. bacterium piece is cut, and size is 3.5mm × 20mm, is used after low temperature sterilization.
2. spore solution is diluted according to first step bacterium amount count results, concentration is 1.0 × 108Cfu/mL, every bacterium piece drop
Dye 10ul, spontaneously dries overnight or more than 12h.
3. dried bacterium piece is put in Bluepoint curved neck easy breaking ampule bottle, is carried out warm high and is melted envelope.
(3) recovery media is prepared
Quality volume (g/mL) percentage of hot air sterilization biological indicator recovery media is：
Tryptone 2%
Soy peptone 1.5%
Sodium chloride 2.5%
Distilled water is prepared, well mixed to be completely dissolved, and adjusts pH7.2, is used after being sterilized through 121 DEG C.
Recovery media after sterilizing is sub-packed in aseptic plastic pipe, often addition 1.4mL in pipe, screws bottle cap, prevented
Only liquid spills, it is to avoid living contaminants.
(4) finished product assembling, external packing production.
Embodiment 3
(1) bacillus subtilis black variety gemma culture
1. nutrient broth medium 50mL is prepared using 250ml conical flasks, used after 121 DEG C of sterilizings, choose hay bacillus
Var. niger spores 20ul is activated, soak time 5h.
2. medicine is weighed, bacillus subtilis black variety gemma growth medium is prepared, 250mL is dispensed in each gemma bottle,
Room temperature is used after being cooled to solid-state after 121 DEG C of sterilizings.
3. operated in Biohazard Safety Equipment, the bacillus subtilis black variety gemma after activation is mixed, in each gemma bottle
Inoculation 10mL, mixes and swings uniform, after inoculation liquid is fully contacted growth media surface, pours out inoculation liquid.
4. gemma bottle is put in 37 DEG C of bacteriological incubators and cultivates 48h.
5. culture receives bacterium after terminating, and 15mL sterilized waters are added in each gemma bottle, and gemma growth medium is scraped using L rods
The gemma on surface, is poured into aseptic conical flask, is repeated 3 times.
6. the spore solution being collected into carries out 3000 and leaves heart 25min, is repeated 4 times, and centrifugation is cleaned, and uses aseptic water dissolves
The gemma precipitation for finally giving.
7. bacterium amount calculating is carried out to final spore solution with dilution method, be put in 4 DEG C of Refrigerator stores.
(2) bacillus subtilis black variety gemma bacterium piece makes
1. bacterium piece is cut, and size is 3.5mm × 20mm, is used after low temperature sterilization.
2. spore solution is diluted according to first step bacterium amount count results, concentration is 1.0 × 108Cfu/mL, every bacterium piece drop
Dye 10ul, spontaneously dries overnight or more than 12h.
3. dried bacterium piece is put in Bluepoint curved neck easy breaking ampule bottle, is carried out warm high and is melted envelope.
(3) recovery media is prepared
Quality volume (g/mL) percentage of hot air sterilization biological indicator recovery media is：
Tryptone 1%
Soy peptone 0.5%
Sodium chloride 1.5%
Distilled water is prepared, well mixed to be completely dissolved, and adjusts pH7.2, is used after being sterilized through 121 DEG C.
Recovery media after sterilizing is sub-packed in aseptic plastic pipe, often addition 1.4mL in pipe, screws bottle cap, prevented
Only liquid spills, it is to avoid living contaminants.
(4) finished product assembling, external packing production.
When hot air sterilization biological indicator of the present invention is used, will be taken out by the ampoule bottle of sterilizing program, room temperature
After cooling, operated in Biohazard Safety Equipment, ampoule bottle is opened using supporting Medical abrasive wheel, bacterium piece is aseptically shifted
To in the plastic tube of recovery media, result is observed after 37 DEG C of culture 48h.By whether producing precipitation to judge to go out in plastic tube
Bacterium effect.Application method is simple and convenient, is as a result easy to observation.
Claims (10)
1. a kind of hot air sterilization biological indicator, it is characterised in that have the ampoule bottle of bacterium piece and modeling equipped with recovery media by envelope
Expects pipe is constituted, and the two is individually present.
2. hot air sterilization biological indicator according to claim 1, it is characterised in that described bacterium piece is by indicating micro- life
Thing and bacterial carrier are constituted.
3. hot air sterilization biological indicator according to claim 1, it is characterised in that described indicator microoraganism is withered grass
Bacillus var. niger spores.
4. hot air sterilization biological indicator according to claim 1, it is characterised in that described bacterial carrier is glass fibers
GF/F grades of filter paper of dimension.
5. hot air sterilization biological indicator according to claim 1, it is characterised in that described ampoule bottle is Bluepoint song neck
Easy break ampoule bottle.
6. hot air sterilization biological indicator according to claim 1, it is characterised in that the composition of described recovery media
As follows, with quality volume percentage, in terms of g, volume is in terms of mL for quality：
7. hot air sterilization biological indicator according to claim 1, it is characterised in that the preparation of described recovery media
Method is that tryptone, soy peptone and sodium chloride are added in distilled water, well mixed to be completely dissolved, and adjusts pH7.2,
After being sterilized through 121 DEG C, obtain final product.
8. hot air sterilization biological indicator according to claim 1, it is characterised in that described plastic tube is PET freezings
Pipe.
9. the preparation method of any described hot air sterilization biological indicators of a kind of claim 1-8, it is characterised in that step is such as
Under：
(1) indicator microoraganism culture, spore solution is collected；
(2) dye spore solution is dripped on bacterial carrier, dried for standby obtains bacterium piece；
(3) bacterium piece is put in ampoule interior, carries out heat and melt envelope；
(4) recovery media is weighed and prepared；
(5) after recovery media sterilizing, operated in Biohazard Safety Equipment, be sub-packed in plastic tube, screw bottle cap；
(6) finished product assembling, external packing production.
10. the preparation method of hot air sterilization biological indicator according to claim 9, it is characterised in that institute in step (2)
Bacterial content is 1 × 10 in the bacterium piece stated6Cfu/ pieces.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109266554A (en) * | 2018-10-09 | 2019-01-25 | 南京巨鲨显示科技有限公司 | A kind of biological indicator bacterium piece preparation method |
| CN110055299A (en) * | 2019-01-21 | 2019-07-26 | 中山大学 | Biological indicator and preparation method thereof for the instruction that sterilizes |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4291122A (en) * | 1980-08-14 | 1981-09-22 | American Sterilizer Company | Biological indicator for sterilization processes |
| CN102041292A (en) * | 2010-11-04 | 2011-05-04 | 河北省农林科学院遗传生理研究所 | Atmospheric steam sterilizing biological indicator and preparation method thereof |
| CN105087361A (en) * | 2015-09-08 | 2015-11-25 | 湖州一控医疗科技有限公司 | Suspension type biological indicator and preparation method thereof |
-
2016
- 2016-12-09 CN CN201611127865.XA patent/CN106755280A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4291122A (en) * | 1980-08-14 | 1981-09-22 | American Sterilizer Company | Biological indicator for sterilization processes |
| CN102041292A (en) * | 2010-11-04 | 2011-05-04 | 河北省农林科学院遗传生理研究所 | Atmospheric steam sterilizing biological indicator and preparation method thereof |
| CN105087361A (en) * | 2015-09-08 | 2015-11-25 | 湖州一控医疗科技有限公司 | Suspension type biological indicator and preparation method thereof |
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| Title |
|---|
| NORCROSS NL等: "Rapid heat treatment of bacteria. I. Sterilization of suspensions of Serratia marcescens and spores of Bacillus subtilis var. niger.", 《APPL MICROBIOL》 * |
| 张爱萍等: "《药品GMP指南 无菌药品》", 31 August 2011, 中国医药科技出版社 * |
| 朱杏花等: "自含式生物指示剂在干热灭菌效果监测中的研究", 《中华医院感染学杂志》 * |
| 杨华明等: "《现代医院消毒学 第3版》", 28 February 2013, 人民军医出版社 * |
| 黄秀清等: "《杭州湾入海污染物总量控制和减排技术研究》", 31 October 2015, 海洋出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109266554A (en) * | 2018-10-09 | 2019-01-25 | 南京巨鲨显示科技有限公司 | A kind of biological indicator bacterium piece preparation method |
| CN110055299A (en) * | 2019-01-21 | 2019-07-26 | 中山大学 | Biological indicator and preparation method thereof for the instruction that sterilizes |
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Application publication date: 20170531 |