CN1049029A - Biological indicator for sterilization monitor and production method - Google Patents
Biological indicator for sterilization monitor and production method Download PDFInfo
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- CN1049029A CN1049029A CN 90100755 CN90100755A CN1049029A CN 1049029 A CN1049029 A CN 1049029A CN 90100755 CN90100755 CN 90100755 CN 90100755 A CN90100755 A CN 90100755A CN 1049029 A CN1049029 A CN 1049029A
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- mycoderm
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Abstract
The present invention relates to a kind of biological indicator of in sterilization monitor, using and production method thereof, its objective is in order accurately to judge the true effect of sterilization, its main technical schemes is that sterilization indicator liquid is added in a kind of dissolvable matrix, then this mixed solution is dripped on sterile transparent plastic paper or overlay, drying is made mycoderm under proper temperature and humidity, is used to monitor the effect of sterilization.
Description
The present invention relates to a kind of biological indicator of in sterilization monitor, using and production method thereof.
Since the nearly century, with the existing very big development of biology method measuring sterilisation effect, the effect of biological indicator in sterilization quality control is day by day for having emphasized the importance of use biological indicator in the aseptic curable product laws and regulations on the management in the world that people pay attention to, American Pharmacopeia was chapters and sections with " biological monitor " in 1985 separately in the 21st edition, at length narrated and list the biological scraps of paper standard of " biological monitor ".The Soviet Union uses microbiological contamination cloth sheet, Japan to be the indicator sheet with microbiological contamination filter paper, and West Germany's hygiology and microbiology association recommend to be the carrier that carries disease germs with cotton.China introduces microbiological contamination cloth sheet from the fifties from the Soviet Union and uses till today always.But both at home and abroad microbiological contamination cloth sheet that adopts and microbiological contamination filter paper because will sterilize that indicator liquid directly droplet dyes, dip-dye or spray-painting be on cloth sheet or filter paper, mostly there is following shortcoming: because the carrier surface different in kind, even commaterial, the difference of its microbiological contamination amount is also very big, and in the remaining viable bacteria test experience after disinfecting, because carrier is insoluble, the bacterium of staying in the material slit can not all discharge, influence the viable bacteria rate of recovery, cause the instability of experimental result, poor repeatability.Therefore be difficult for reaching stdn, be difficult to medical device disinfection and the strictness of medical treatment product aseptic quality are controlled, had a strong impact on the progress of China's sterilization scientific effort.We think for this reason: setting up a standardized new bio indicator is the key point of development China sterilization cause.
The objective of the invention is to avoid the weak point in the above-mentioned technology and a kind of standard, easy to use, good solubility biological indicator and the production method thereof of repeatability be provided.
Purpose of the present invention can reach by following measure: we adopt Staphylococcus albus (Staphylococcus albus 8032), streptococcus aureus (Staphylococcus aureus ATCC 6538), streptococcus faecium (Streptococcus faecium ATCC 19581), spores of bacillus subtilin black mutation (B.subtilis var.niger ATCC 9372), bacillus pumilis gemma (B.pumilus E601), wax shape bacillus gemma (B.cereus SSI, C the sterilization indicator
1/ 1), bacillus stearothermophilus (B.stearothermophilus SSI, K31).Four kinds of genus bacillus are inoculated in TGY agar (to be contained
Peptone 0.5%, racemization glucose 0.1%, yeast extract 0.3%, agar 1.8%), remove bacstearothermophilus and be incubated at 56 ℃, 3-5 beyond the highest heavens, its excess-three kind gemma was all cultivated 2-5 days down at 37 ℃, washed lawn with distilled water, gave a baby a bath on the third day after its birth altogether time, preserved in 4 ℃ of refrigerators.Three kinds of bacterial propagules were all cultivated 24 hours down at 37 ℃ with plain agar, and lawn washes with the 0.03M phosphate buffered saline buffer, and it is standby to put in 4 ℃ of refrigerators preservation, gemma and propagulum be the 4th generation culture.
Bacterium liquid is added mixing in the dissolvable matrix, and wherein dissolvable matrix is by 0.5%-2% gelatin, 0.5%-2% polyoxyethylene glycol, 0.5%-1% glycerol and the preparation of 0.03M phosphate buffered saline buffer, then with 10
5-10
6Individual bacterium is one to drip on sterile transparent plastic paper or overlay, and the amount that preferably makes every is 45-55 μ l, is that 18-28 ℃, humidity are under the condition of 50%-70% after the drying in temperature then, with the plastic film sealing or be collected in the vial.
Be the test that stability, characteristics etc. to this kind biological indicator (abbreviation mycoderm) are carried out below.Make live bacterial count with 10 times of dilution methods of successively decreasing, every mycoderm bacteria containing amount is represented with colony-forming unit (CFU)/sheet.
One. the difference of bacteria containing amount between mycoderm: containing bacterial propagule with the mycoderm of dissolvable matrix preparation is 8.4 * 10
5-8.0 * 10
6The CFU/ sheet; Gemma is 4.4 * 10-5.1 * 10CFU/ sheet.
With the Staphylococcus albus mycoderm is that example is checked 159 of mycoderms altogether through 29 tests, comprises the error of operative technique, and the specific inaccuracy of viable count is accounting for 87.9% of inspection mycoderm sum between every below 10%.
Two. the recovery of mycoderm viable bacteria: mycoderm liquid that is mixed with in proportion and dried mycoderm carry out viable count respectively and relatively reach the t test, prove that system film and drying process are little to above several bacteria living influences, difference that there are no significant.Except that borrowing the shape bacillus gemma rate of recovery low slightly (65.86%), all the other various mycoderm rate of recovery are all more than 80%, and high person reaches 98.65%.
Three. the stability of mycoderm: to being stored in the Staphylococcus albus mycoderm of room temperature (18-20 ℃) and refrigerator (4 ℃), do the observation that live bacterial count carried out for 13 weeks altogether weekly, the mycoderm of preserving in the proof refrigerator is more stable than room temperature preservation, and the viable count that lasts after 3 months in the every mycoderm still remains on 10
6More than the CFU.Streptococcus aureus in 4 ℃ down keep 3 months after the viable bacteria rate slightly have a declining tendency, gemma remains in that viable count does not also have obvious variation in the refrigerator.
The stability of 5 kinds of mycoderms relatively
| Bacterial classification | Shelf time (week) | Every mycoderm contains viable count (CFU/ sheet) | |
| During preparation (XSD) | Preserve back (XSD) | ||
| Staphylococcus albus | 13 | 7.73×10 6±0.55 | 1.51×10 6±0.3 |
| Streptococcus aureus | 12 | 1.17×10 6±0.15 | 1.04×10 6±0.15 |
| The mutation of spores of bacillus subtilin black | 25 | 5.13×10 6±0.15 | 1.72×10 6±0.51 |
| The bacillus pumilis gemma | 8 | 5.47×10 6±0.5 | 4.13×10 6±1.02 |
| Wax shape bacillus gemma | 8 | 3.03×10 6±1.1 | 1.83×10 6±0.72 |
Streptococcus aureus mycoderm its resistibility under the effect of 7500ppm phenylic acid is not less than the bacterium liquid before the system film.Mycoderm and bacterium liquid is stable identical in 60 ℃ of environment.The resistibility of mycoderm through preserving Surchlor GR 60 all around with bacillus subtilis spore black mutation preparation is 7500ppm, 30 minutes.The streptococcus aureus mycoderm needs 24-26 minute to the 7500ppm phenylic acid, and its drags are constant after five weeks of preservation.
With 1 * 10
6CFU/ml Staphylococcus albus bacterium liquid is pressed 0.01ml/ sheet microbiological contamination amount, and " test organisms cloth " live bacterial count of 0.5 * 1.0cm size of preparation is 3 * 10
3The CFU/ sheet, after one week, every is 0.7 * 10 in room temperature preservation
2CFU, refrigerator preserver are 1.2 * 10
2CFU has viable bacteria decline or the loss of 96%-98%.
Four. the application of mycoderm on sterilization monitor: we have also carried out monitoring aspect following to various mycoderms except that the qualitative monitoring that is used for the chemical liquid sterilizing agent.
1. to the monitoring of disinfection by ultraviolet light: measure and regulate irradiation distance to required exposure intensity with UVR-254 type ultraviolet ray intensity meter, place aseptic plate to be exposed to mycoderm and calculate its killing rate under the different exposure intensities.
2. to the monitoring of microwave disinfection: mycoderm is put into 16 * 16 * 8cm size dressing bag middle part respectively, external application wet towel parcel, in output rating is under the 650W microwave action, Staphylococcus albus, streptococcus aureus and streptococcus faecium are dead in 4 minutes, and the thermophilic bacteria gemma needs could all kill after the effect in 10 minutes.
3. to the monitoring of high pressure steam sterilization: meet water because of mycoderm and promptly dissolve, when high pressure steam sterilization, mycoderm is placed on respectively in the aseptic empty test tube, place pre-pasteurized article middle part, after the sterilization liquid nutrient medium poured in the mycoderm pipe and cultivate then, the result proves that 15 pounds were killed Staphylococcus albus, streptococcus aureus, streptococcus faecium, spores of bacillus subtilin, wax shape bacillus gemma and bacillus pumilis gemma in 20 minutes, and bacillus stearothermophilus need be increased to 20 pounds or extend to the deactivation of 30 minutes ability.
4. to the monitoring of gaseous decontaminant sterilisation effect: mycoderm is placed aseptic plate respectively, is 0.15m at volume
3Carry out gas depoisoning test (20 ℃ of room temperatures in the dry cylinder of wooden case and 9 liters, relative humidity 80%) select 10% glutaraldehyde for use, dosage 1.06ml/ liter acts on that three kinds of gemma to Staphylococcus albus, streptococcus faecium and wax shape bacillus, bacillus pumilis, bacillus stearothermophilus have killing action after 40 hours.Through effect in 22 hours, except that above five kinds of bacterium cyclophanes being grown body and gemma has the killing action, and streptococcus aureus and spores of bacillus subtilin also there is deactivation with 0.05ml/ liter formalin.We also use 26 gram/m
3Acid chlorine is made the sootiness sterilizing test, can reach 77% after acting on 30 minutes to Staphylococcus albus and killing 32%, 1 hour.
Make biological indicator according to above-mentioned experimental result proof with the mycoderm of dissolvable matrix preparation, the sterilisation effect mensuration of ultraviolet ray, microwave, high pressure steam and gaseous decontaminant is suitable for.
The present invention has following advantage compared to existing technology: but but 1. good reproducibility 4. carriers of high 3. tests of constant 2. rate of recovery of bacterium number to low 5. prolonged preservation of sterilizing agent rate of consumption 6. 7. long-distance transports free from environmental pollution 8. 9. stdn mass production with low cost easy to use.
The present invention will now be further detailed embodiment: streptococcus aureus (staphylococcus aureus ATCC 6538) was cultivated 24 hours down at 37 ℃ with plain agar, lawn washes with the 0.03M phosphate buffered saline buffer, put preserve in 4 ℃ of refrigerators standby.
With 0.03M phosphate buffered saline buffer preparation 1 * 10
9The bacterium liquid of individual/ml is with dissolvable matrix (gelatin 1%, polyoxyethylene glycol 1%, glycerol 0.5%, 0.03M phosphate buffered saline buffer) 100ml drips on aseptic transparent plastic film with 50 microlitres one with after 2ml bacterium liquid mixes, 25 ℃, dry under the condition of RH50%, seal with plastic film.
Claims (8)
1, a kind of biological indicator for sterilization monitor is characterized in that it is the mycoderm of being made by sterilization indicator and dissolvable matrix, and dissolvable matrix comprises gelatin, polyoxyethylene glycol and glycerol.
2, biological indicator according to claim 1 is characterized in that each mycoderm contains 10
5-10
6Individual indicator.
3, biological indicator according to claim 1 is characterized in that dissolvable matrix is made up of 0.5-2% gelatin, 0.5-2% polyoxyethylene glycol, 0.5-1% glycerol and 0.03M phosphate buffered saline buffer.
4, biological indicator according to claim 1 is characterized in that above-mentioned mycoderm is sealed in transparent plastic paper or the overlay.
5, a kind of production method of biological indicator for sterilization monitor, comprise after the cultivation of sterilization indicator, be made into bacterium liquid with 0.03M phosphate buffered saline buffer or distilled water, it is characterized in that above-mentioned bacterium liquid is added in the dissolvable matrix, dissolvable matrix is made up of 0.5-2% gelatin, 0.5-2% polyoxyethylene glycol, 0.5-1% glycerol and 0.03M phosphate buffered saline buffer, then above-mentioned mixed solution is dripped on sterile transparent plastic paper or overlay, seal after being dried to mycoderm.
6, production method according to claim 5 is characterized in that mycoderm is an exsiccant under temperature 18-28 ℃, the condition of humidity 50-70%.
7, a kind of production method as claimed in claim 5, each bacteria containing amount that drips is 10 when it is characterized in that dripping solubility indicator liquid
5-10
6Individual.
8, a kind of production method as claimed in claim 5, the amount that it is characterized in that each are 45-55 μ l.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 90100755 CN1049029A (en) | 1990-02-19 | 1990-02-19 | Biological indicator for sterilization monitor and production method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 90100755 CN1049029A (en) | 1990-02-19 | 1990-02-19 | Biological indicator for sterilization monitor and production method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1049029A true CN1049029A (en) | 1991-02-06 |
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ID=4876791
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 90100755 Pending CN1049029A (en) | 1990-02-19 | 1990-02-19 | Biological indicator for sterilization monitor and production method |
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| CN (1) | CN1049029A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105779563A (en) * | 2016-04-29 | 2016-07-20 | 大连华立金港药业有限公司 | Detection reagent kit and method of heat-resistant microorganisms in elemene lipidosome injection semi-finished products |
| CN108818863A (en) * | 2018-05-30 | 2018-11-16 | 湖州汇德集团有限公司 | A kind of drying anticorrosion process of solid wooden floor board |
-
1990
- 1990-02-19 CN CN 90100755 patent/CN1049029A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105779563A (en) * | 2016-04-29 | 2016-07-20 | 大连华立金港药业有限公司 | Detection reagent kit and method of heat-resistant microorganisms in elemene lipidosome injection semi-finished products |
| CN108818863A (en) * | 2018-05-30 | 2018-11-16 | 湖州汇德集团有限公司 | A kind of drying anticorrosion process of solid wooden floor board |
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