CN110055299A - Biological indicator and preparation method thereof for the instruction that sterilizes - Google Patents
Biological indicator and preparation method thereof for the instruction that sterilizes Download PDFInfo
- Publication number
- CN110055299A CN110055299A CN201910056608.9A CN201910056608A CN110055299A CN 110055299 A CN110055299 A CN 110055299A CN 201910056608 A CN201910056608 A CN 201910056608A CN 110055299 A CN110055299 A CN 110055299A
- Authority
- CN
- China
- Prior art keywords
- culture medium
- preparation
- sterilizes
- biological indicator
- instruction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000090 biomarker Substances 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 230000001954 sterilising effect Effects 0.000 claims abstract description 28
- 239000001963 growth medium Substances 0.000 claims abstract description 27
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 24
- 241000894006 Bacteria Species 0.000 claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000012153 distilled water Substances 0.000 claims abstract description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 10
- 239000011521 glass Substances 0.000 claims abstract description 10
- 241000193385 Geobacillus stearothermophilus Species 0.000 claims abstract description 9
- XJRPTMORGOIMMI-UHFFFAOYSA-N ethyl 2-amino-4-(trifluoromethyl)-1,3-thiazole-5-carboxylate Chemical compound CCOC(=O)C=1SC(N)=NC=1C(F)(F)F XJRPTMORGOIMMI-UHFFFAOYSA-N 0.000 claims abstract description 7
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims abstract description 6
- 230000001580 bacterial effect Effects 0.000 claims abstract description 6
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000012137 tryptone Substances 0.000 claims abstract description 6
- 239000011780 sodium chloride Substances 0.000 claims abstract description 5
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims abstract description 4
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000004474 valine Substances 0.000 claims abstract description 4
- 229940041514 candida albicans extract Drugs 0.000 claims abstract description 3
- 238000004132 cross linking Methods 0.000 claims abstract description 3
- 239000003365 glass fiber Substances 0.000 claims abstract description 3
- 239000012138 yeast extract Substances 0.000 claims abstract description 3
- 241000726221 Gemma Species 0.000 claims description 17
- 238000011084 recovery Methods 0.000 claims description 13
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 10
- 239000003708 ampul Substances 0.000 claims description 9
- 238000004090 dissolution Methods 0.000 claims description 8
- 239000012452 mother liquor Substances 0.000 claims description 4
- 238000012856 packing Methods 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 239000007850 fluorescent dye Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- UPSFMJHZUCSEHU-JYGUBCOQSA-N n-[(2s,3r,4r,5s,6r)-2-[(2r,3s,4r,5r,6s)-5-acetamido-4-hydroxy-2-(hydroxymethyl)-6-(4-methyl-2-oxochromen-7-yl)oxyoxan-3-yl]oxy-4,5-dihydroxy-6-(hydroxymethyl)oxan-3-yl]acetamide Chemical compound CC(=O)N[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@H](O)[C@@H](NC(C)=O)[C@H](OC=2C=C3OC(=O)C=C(C)C3=CC=2)O[C@@H]1CO UPSFMJHZUCSEHU-JYGUBCOQSA-N 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 2
- 229910021529 ammonia Inorganic materials 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- 238000009423 ventilation Methods 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 21
- 239000012528 membrane Substances 0.000 abstract description 3
- 230000000249 desinfective effect Effects 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 15
- ARQXEQLMMNGFDU-JHZZJYKESA-N 4-methylumbelliferone beta-D-glucuronide Chemical compound C1=CC=2C(C)=CC(=O)OC=2C=C1O[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O ARQXEQLMMNGFDU-JHZZJYKESA-N 0.000 description 7
- ARQXEQLMMNGFDU-UHFFFAOYSA-N 4MUG Natural products C1=CC=2C(C)=CC(=O)OC=2C=C1OC1OC(C(O)=O)C(O)C(O)C1O ARQXEQLMMNGFDU-UHFFFAOYSA-N 0.000 description 7
- 238000005119 centrifugation Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000010170 biological method Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- MBHINSULENHCMF-UHFFFAOYSA-N n,n-dimethylpropanamide Chemical compound CCC(=O)N(C)C MBHINSULENHCMF-UHFFFAOYSA-N 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 108010023063 Bacto-peptone Proteins 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- YUDPTGPSBJVHCN-JZYAIQKZSA-N 4-Methylumbelliferyl-alpha-D-glucopyranoside Chemical compound C1=CC=2C(C)=CC(=O)OC=2C=C1O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YUDPTGPSBJVHCN-JZYAIQKZSA-N 0.000 description 1
- 238000009631 Broth culture Methods 0.000 description 1
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N D-alpha-Ala Natural products CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 description 1
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- QNAYBMKLOCPYGJ-UWTATZPHSA-N L-Alanine Natural products C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 229960003767 alanine Drugs 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000000721 bacterilogical effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- -1 bromine cresols Chemical class 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 238000002845 discoloration Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- 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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Physics & Mathematics (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Toxicology (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention is a kind of biological indicator and preparation method thereof for the instruction that sterilizes, and is detected to the validity of high pressure steam sterilization process.Sterile biological indicator is made of the plastic tube for being packaged with bacterium piece with the glass tube for restoring culture medium is packaged with, and the two is individually present before use, is not interfere with each other.Bacterium piece is made of indicator microoraganism and bacterial carrier, indicator microoraganism is Geobacillus stearothermophilus, bacterial carrier is glass fiber filter paper, restoring culture medium to form plastic tube by tryptone, soy peptone, yeast extract, sodium chloride, bromocresol purple, 4-methyl umbelliferone-α-D- glucopyranoside, l-Alanine, Valine, distilled water etc. is the translucent high temperature resistant type cross-linking plastic pipe that top is provided with venthole, and ventilated membrane is covered, so that disinfecting process mesohigh steam is come into full contact with bacterium piece.The advantages that simple with preparation, raw material is easy to get, as a result reliable.
Description
Technical field
The invention belongs to biological monitoring technical fields in sterilization, and in particular to a kind of to refer to for the biology indicated that sterilizes
Show agent and preparation method thereof.
Background technique
With the progress of medical technique level, new medical instrument and medical supplies are continuously emerged.In hospital's routine work
In, the sterilizing of medical instrument is an important content.The validity for wherein how detecting sterilization of medical instrument keeps its detection qualified
After can efficiently hospital circulation using be even more the most important thing.The sterilizing methods that modern hospital uses usually have high pressure steam sterilization
Disinfection, oxirane disinfection, radiation sterilization etc., wherein high pressure steam sterilization disinfection is most widely used, and being suitable for can high temperature resistant
Metal scalpel cut equal medical instruments, sterilization effect is good, can kill all microorganisms (including gemma) in bacterium.High pressure is steamed
The monitoring method of vapour sterilizing generally comprises chemical method and biological method, and chemical method is usually made of chemical alternating temperature ink, uses
Whether experienced high pressure-temperature in the apparatus for indicating subject to sterilization, belongs to process indicator, this method is using simply, and cost is relatively low.
It is widely used in the process monitoring of sterilization process, but the shortcomings that chemical finger-length measurement is that medical instrument can only be prompted to experienced height
The process of super pressure-high temperature, can not indicate whether final microorganism is killed, therefore biological method is still in Disinfection examination
" Jin Zhibiao ".Biological method develops by several generations, first generation culture medium cultivation, by directly observing in culture medium whether have bacterium
It is born into determine sterilization effect.This method usually requires time-consuming 7 days, and the operation for hospital is very unfavorable.Second on behalf of
Self-contained biological indicator, by bacterium piece and culture set of bases to one, generating acid using gemma recovery declines medium pH
The method of indicator discoloration is set to indicate sterilization effect, this method is more simple and convenient for use departing from laboratory cultures base culture,
Incubation time is substantially reduced to 24-48h, but the time is still long, is not able to satisfy the requirement of modern hospital.Therefore, develop
A kind of biological indicator that the used time is shorter is even more important.
Summary of the invention
The object of the present invention is to provide a kind of new sterile biological indicator, using easy, result is reliable, to sterilization process
Validity detected；It is a further object of the present invention to provide the preparation method of sterile biological indicator, economical and efficient is easy to
Operation.
To achieve the above object, sterile biological indicator of the present invention is plastic tube and the encapsulation by being packaged with bacterium piece
It is made of the glass tube for restoring culture medium, the two is individually present before use, is not interfere with each other.
The bacterium piece is made of indicator microoraganism and bacterial carrier.The indicator microoraganism for stearothermophilus bud
Spore bacillus (Bacillus stearothermophilus, ATCC 7953).The bacterial carrier be glass fiber filter paper or
Other carriers.
The vial is ampoule bottle.Ampoule bottle, which need to melt envelope through overheat, keeps both ends silent, it is ensured that bacterium piece is using preceding place
In dry gnotobasis.
The plastic tube is that top is provided with the translucent high temperature resistant type cross-linking plastic pipe of venthole, and covers ventilated membrane, is made
Disinfecting process mesohigh steam can come into full contact with bacterium piece.
The recovery culture medium is by tryptone, soy peptone, yeast extract, sodium chloride (NaCl), bromine cresols
Purple, 4-methyl umbelliferone-α-D- glucopyranoside (4-Methylumbelliferyl- α-D-glucopyranoside, 4-
MUG), l-Alanine, Valine, distilled water etc. form.
The preparation method of the recovery culture medium mother liquor is that tryptone, soy peptone etc. are added to distilled water
In, it is uniformly mixed after being completely dissolved and bromocresol purple is added, pH to 7.0-7.5 is adjusted, to get mother liquor after high-temp steam sterilizing.
L-Alanine is dissolved with partial mother liquid, with n,N-Dimethylformamide (N, N-Dimethylformamide, DMF) or other conjunctions
After suitable solvent dissolution fluorogenic substrate 4-MUG, packing is into glass ampoule bottles in clean environment.
The preparation method of high pressure steam sterilization biological indicator of the present invention, the specific steps are as follows:
(1) Geobacillus stearothermophilus is cultivated, gemma is prepared, and the gemma piece containing specified gemma number is made；
(2) bacterium piece is fixed on to the lower section of sterilizing plastic tube；
(3) preparation of culture medium is restored；
(4) after the culture medium sterilization prepared, glass ampule pipe is encapsulated into clean environment；
(5) by after ampoul tube and plastic tube assembling, upper tube cap is pressed；
(6) finished product assembles, outer packing production.
Spore content is 1 × 10 in bacterium piece described in step (2)5-1×106Cfu/ piece.
Compared with the prior art, the present invention has the following beneficial effects:
(1) it is detected using the present invention for the rapid fluorescence of high pressure steam sterilization process, has preparation simple, raw material is easy
, it is as a result reliable the advantages that, can be used for the quick detection of high-pressure steam sterilizing result.
(2) the biological indicator system contained by the present invention is individually present mutually, is being in dry sterile shape using preceding bacterium piece
State, gemma state keep stablizing, and are stabilized product in shelf-life and transportational process.
(3) steam can be come into full contact with the present invention by gland aperture and bacterium piece in use.Make biological indicator system
System can good participation sterilization process, and the report of gemma anabiosis rate is provided after the completion of sterilizing by fluorescence detection equipment
It accuses.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of the biological indicator.
In Fig. 1, (1) aperture plastic cap, (2) bacteriological filtration ventilated membrane, (3) are ampoul tube, and (4) plastic outer tube, (5) are thermophilic rouge
Fat bacillus spore bacterium piece.
Specific embodiment
Below by specific embodiment, the present invention is further explained, and following embodiment facilitates those skilled in the art
The present invention is further understood, but is not intended to limit protection scope of the present invention.
The preparation of the recovery culture medium of embodiment 1
Quality volume (g/100mL) percentage of biological indicator recovery culture medium are as follows:
Step: (1) weighing above-mentioned appropriate bactopeptone and LB culture medium, and after the dissolution of appropriate distilled water is added, adds
Distilled water constant volume is added as 1L after entering bromocresol purple.By this basic culture solution after disinfection with high pressure steam sterilizes, it is cooled to room
Temperature.
(2) dissolution is sufficiently stirred after l-Alanine being added in above-mentioned culture solution.
(3) pH value of solution is adjusted to 7.0-7.4 with appropriate KOH.
(4) in clean environment, with appropriate N, N- dimethylpropionamide dissolves 4-MUG.
(5) in clean environment, dissolved 4-MUG is added in basic culture solution, is sufficiently stirred to obtain recovery culture
Base.
(6) in clean environment, sealing after culture medium is distributed into glass ampule will be restored, every pipe is packed into 0.7mL.
2 bacillus stearothermophilus gemma culture of embodiment
Step: (1) it using 250ml conical flask prepares pancreas peptone soybean broth culture medium 50mL, makes after 121 DEG C of sterilizings
With selection bacillus stearothermophilus 20ul, in 60 DEG C of amplification cultivations.
(2) bacillus stearothermophilus gemma growth medium is prepared, room temperature makes after being cooled to solid-state after 121 DEG C of sterilizings
With.
(3) it is operated in Biohazard Safety Equipment, bacillus stearothermophilus is inoculated in each production spore plate respectively, will be produced
Spore flat-plate inverted, which is postponed, cultivates 72h at 60 DEG C.
(5) spore solution being collected into carries out 4000 turns of centrifugation 20min, is repeated 4 times, and centrifugation is cleaned, using sterile water-soluble
Solve finally obtained gemma precipitating.
(4) gemma is collected after cultivating, using the gemma of L stick scraping gemma growth media surface, is distilled using ice
Topple over after water washing in sterile conical flasks, is repeated 3 times.
(5) spore solution being collected into carries out 4000 turns of centrifugation 20min, is repeated 4 times, and centrifugation is cleaned, using sterile water-soluble
Solve finally obtained gemma precipitating.
(6) bacterium amount calculating is carried out to final spore solution with dilution method, is put in 4 DEG C of refrigerators and saves.
The preparation of the recovery culture medium of embodiment 3
Quality volume (g/100mL) percentage of biological indicator recovery culture medium are as follows:
Step: (1) weighing above-mentioned appropriate bactopeptone and LB culture medium, and after the dissolution of appropriate distilled water is added, adds
Distilled water constant volume is added as 1L after entering bromocresol purple.By this basic culture solution after disinfection with high pressure steam sterilizes, it is cooled to room
Temperature.
(2) dissolution is sufficiently stirred after Valine being added in above-mentioned culture solution.
(3) pH value of solution is adjusted to 7.0-7.4 with KOH.
(4) in clean environment, with appropriate N, N- dimethylpropionamide dissolves 4-MUG.
(5) in clean environment, dissolved 4-MUG is added in basic culture solution, is sufficiently stirred to obtain recovery culture
Base.
(6) in clean environment, sealing after culture medium is distributed into glass ampule will be restored, every pipe is packed into 0.7mL.
The preparation of the recovery culture medium of embodiment 4
Quality volume (g/100mL) percentage of biological indicator recovery culture medium are as follows:
Step: (1) weighing above-mentioned appropriate tryptone, soy peptone and NaCl, and after the dissolution of appropriate distilled water is added,
Distilled water constant volume is added as 1L after bromocresol purple is added.By this basic culture solution after disinfection with high pressure steam sterilizes, it is cooled to
Room temperature.
(2) dissolution is sufficiently stirred after l-Alanine being added in above-mentioned culture solution.
(3) pH value of solution is adjusted to 7.0-7.4 with KOH.
(4) in clean environment, with appropriate N, N- dimethylpropionamide dissolves 4-MUG.
(5) in clean environment, dissolved 4-MUG is added in basic culture solution, is sufficiently stirred to obtain recovery culture
Base.
(6) in clean environment, sealing after culture medium is distributed into glass ampule will be restored, every pipe is packed into 0.7mL.
Claims (5)
1. a kind of biological indicator for the instruction that sterilizes, it is characterised in that: by being packaged with the plastic tube of bacterium piece and being packaged with multiple
The glass tube of former culture medium forms；The bacterium piece is made of indicator microoraganism and bacterial carrier, the indicator microoraganism
For Geobacillus stearothermophilus, the bacterial carrier is glass fiber filter paper, and the plastic tube is that top is provided with ventilation
The translucent high temperature resistant type cross-linking plastic pipe in hole.
2. the biological indicator according to claim 1 for the instruction that sterilizes, it is characterised in that: the recovery culture medium
By tryptone, soy peptone, yeast extract, sodium chloride, bromocresol purple, 4-methyl umbelliferone-α-D- glucopyranoside,
The composition such as l-Alanine, Valine, distilled water.
3. the biological indicator according to claim 1 for the instruction that sterilizes, it is characterised in that: spore content is in bacterium piece
1×105-1×106Cfu/ piece.
4. a kind of preparation method of the biological indicator for the instruction that sterilizes, it is characterised in that the following steps are included:
(1) Geobacillus stearothermophilus is cultivated, gemma is prepared, and the gemma piece containing specified gemma number is made；
(2) bacterium piece is fixed on to the lower section of sterilizing plastic tube；
(3) preparation of culture medium is restored；
(4) after the culture medium sterilization prepared, glass ampule pipe is encapsulated into clean environment；
(5) by after ampoul tube and plastic tube assembling, upper tube cap is pressed；
(6) finished product assembles, outer packing production.
5. the preparation method of the biological indicator according to claim 4 for the instruction that sterilizes, it is characterised in that: restore training
The preparation method for supporting base mother liquor is that tryptone, soy peptone etc. are added in distilled water, is uniformly mixed after being completely dissolved
Bromocresol purple is added, pH to 7.0-7.5 is adjusted, to get mother liquor after high-temp steam sterilizing；The third ammonia of L- is dissolved with partial mother liquid
Acid, after n,N-Dimethylformamide or other suitable solvent dissolution fluorogenic substrate 4-methyl umbelliferone-α-D- glucopyranosides,
Packing is into glass ampoule bottles in clean environment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910056608.9A CN110055299A (en) | 2019-01-21 | 2019-01-21 | Biological indicator and preparation method thereof for the instruction that sterilizes |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910056608.9A CN110055299A (en) | 2019-01-21 | 2019-01-21 | Biological indicator and preparation method thereof for the instruction that sterilizes |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110055299A true CN110055299A (en) | 2019-07-26 |
Family
ID=67316405
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910056608.9A Pending CN110055299A (en) | 2019-01-21 | 2019-01-21 | Biological indicator and preparation method thereof for the instruction that sterilizes |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110055299A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110862920A (en) * | 2019-12-02 | 2020-03-06 | 吴权辉 | Container for biological indicator |
| CN110980004A (en) * | 2019-12-02 | 2020-04-10 | 吴权辉 | Biological indicator for sterilization of medical instruments |
| CN111437419A (en) * | 2020-05-18 | 2020-07-24 | 衡水诺盾生物科技有限公司 | Sterilization verification device for pressure steam sterilization extremely-fast comprehensive challenge and reuse dressing |
| CN113398306A (en) * | 2021-06-30 | 2021-09-17 | 山东新华医疗器械股份有限公司 | Rapid biological indicator for sterilization effect monitoring and use method |
| CN113862170A (en) * | 2021-03-29 | 2021-12-31 | 成都医学院 | Biological indicator for monitoring sterilization effect and preparation method thereof |
| CN114410540A (en) * | 2022-02-07 | 2022-04-29 | 山东新华医疗器械股份有限公司 | Bacillus stearothermophilus culture solution capable of improving spore rate and culture method |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090068716A1 (en) * | 2006-04-11 | 2009-03-12 | Ngk Insulators, Ltd. | Biological indicator and method for producing the same |
| CN104212875A (en) * | 2014-09-09 | 2014-12-17 | 四川科伦药业股份有限公司 | Method for measuring initial spore number of bacillus stearothermophilus in biological indicator |
| 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 |
| CN105907632A (en) * | 2016-04-27 | 2016-08-31 | 南京巨鲨显示科技有限公司 | Genetic engineering biological indicator |
| CN106497851A (en) * | 2016-12-20 | 2017-03-15 | 山东新华医疗器械股份有限公司 | Promote the culture medium of bacillus stearothermophilus fast germination |
| CN106794271A (en) * | 2014-10-10 | 2017-05-31 | 3M创新有限公司 | Use the biological sterilization indicator of resistant to the sterilization agent instrumentality |
| CN106755280A (en) * | 2016-12-09 | 2017-05-31 | 山东新华医疗器械股份有限公司 | Hot air sterilization biological indicator and preparation method thereof |
| CN106867940A (en) * | 2017-03-16 | 2017-06-20 | 青岛农业大学 | A kind of method for promoting the growth of bacillus stearothermophilus gemma to sprout |
-
2019
- 2019-01-21 CN CN201910056608.9A patent/CN110055299A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090068716A1 (en) * | 2006-04-11 | 2009-03-12 | Ngk Insulators, Ltd. | Biological indicator and method for producing the same |
| CN104212875A (en) * | 2014-09-09 | 2014-12-17 | 四川科伦药业股份有限公司 | Method for measuring initial spore number of bacillus stearothermophilus in biological indicator |
| CN106794271A (en) * | 2014-10-10 | 2017-05-31 | 3M创新有限公司 | Use the biological sterilization indicator of resistant to the sterilization agent instrumentality |
| 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 |
| CN105907632A (en) * | 2016-04-27 | 2016-08-31 | 南京巨鲨显示科技有限公司 | Genetic engineering biological indicator |
| CN106755280A (en) * | 2016-12-09 | 2017-05-31 | 山东新华医疗器械股份有限公司 | Hot air sterilization biological indicator and preparation method thereof |
| CN106497851A (en) * | 2016-12-20 | 2017-03-15 | 山东新华医疗器械股份有限公司 | Promote the culture medium of bacillus stearothermophilus fast germination |
| CN106867940A (en) * | 2017-03-16 | 2017-06-20 | 青岛农业大学 | A kind of method for promoting the growth of bacillus stearothermophilus gemma to sprout |
Non-Patent Citations (3)
| Title |
|---|
| 姚楚水: "压力蒸汽灭菌生物指示剂", 《中国消毒学杂志》 * |
| 崔学文等: "自含式压力蒸汽灭菌生物指示剂D值测定的影响因素", 《中国消毒学杂志》 * |
| 李俊华等: "发酵罐的生物指示剂挑战试验方法设计", 《广西轻工业》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110862920A (en) * | 2019-12-02 | 2020-03-06 | 吴权辉 | Container for biological indicator |
| CN110980004A (en) * | 2019-12-02 | 2020-04-10 | 吴权辉 | Biological indicator for sterilization of medical instruments |
| CN110862920B (en) * | 2019-12-02 | 2020-08-25 | 山东贝瑞康生物科技有限公司 | Container for biological indicator |
| CN111437419A (en) * | 2020-05-18 | 2020-07-24 | 衡水诺盾生物科技有限公司 | Sterilization verification device for pressure steam sterilization extremely-fast comprehensive challenge and reuse dressing |
| CN113862170A (en) * | 2021-03-29 | 2021-12-31 | 成都医学院 | Biological indicator for monitoring sterilization effect and preparation method thereof |
| CN113862170B (en) * | 2021-03-29 | 2024-04-30 | 成都医学院 | Biological indicator for monitoring sterilization effect and preparation method thereof |
| CN113398306A (en) * | 2021-06-30 | 2021-09-17 | 山东新华医疗器械股份有限公司 | Rapid biological indicator for sterilization effect monitoring and use method |
| CN113398306B (en) * | 2021-06-30 | 2023-06-30 | 山东新华医疗器械股份有限公司 | Extremely fast biological indicator for monitoring sterilization effect and use method |
| CN114410540A (en) * | 2022-02-07 | 2022-04-29 | 山东新华医疗器械股份有限公司 | Bacillus stearothermophilus culture solution capable of improving spore rate and culture method |
| CN114410540B (en) * | 2022-02-07 | 2024-01-19 | 山东新华医疗器械股份有限公司 | Bacillus stearothermophilus culture solution for improving spore rate and culture method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110055299A (en) | Biological indicator and preparation method thereof for the instruction that sterilizes | |
| CN101893589A (en) | Sterility test method and totally closed bacteria collection ampoule incubator used thereby | |
| CN103045529B (en) | Method of inducing phytophthora nicotianae breda de haan bacteria to generate sporangiums and release zoospores | |
| CN206970617U (en) | Simplify the quick bio indicator of structure | |
| CN109880758A (en) | A kind of lactobacillus plantarum mutagenic strain and its method of mutagenesis and application | |
| JP6975701B2 (en) | Bacillus subtilis isolation method, Bacillus subtilis, microbial preparation containing Bacillus subtilis, medium set for Bacillus subtilis isolation | |
| CN106367346A (en) | Mesenchymal stem cell extraction and perfusion culture system | |
| CN104403965B (en) | A kind of roost rose of Sharon pseudomonad of water body tetracycline pollutant of degrading and its application | |
| CN107505311A (en) | The quick method and biological indicator for determining sterilization effect | |
| CN101485300B (en) | Method for cultivating Acrobeloides nanus | |
| CN106399180A (en) | Acetochlor herbicide degrading bacteria and production method and use of agent of acetochlor herbicide degrading bacteria | |
| CN105441350B (en) | Produce the bacillus subtilis with a variety of enzymatic activitys and its application | |
| Darmady et al. | Radiation sterilization | |
| CN215652849U (en) | Rapid biological indicator for monitoring sterilization effect | |
| CN205408770U (en) | Hothouse plants cultivation device | |
| CN106755280A (en) | Hot air sterilization biological indicator and preparation method thereof | |
| CN101591695A (en) | The sodium lauryl sulphate neutralization increases bacterial context soup | |
| CN101353699A (en) | Polyase chain reaction detecting method of Campylobacter jejuni | |
| CN110885873A (en) | Reagent for blood culture bottle production and blood culture bottle production process | |
| CN106086159A (en) | A kind of zymolyte culture medium that can simultaneously detect two kinds of fecal pollution indicator bacterias and application thereof | |
| CN201648404U (en) | Novel two-phase culture medium of bacillus tuberculosis typus bovinus | |
| CN105628663B (en) | The method for quickly detecting of high-moisture food sterilization effect | |
| CN110607243A (en) | Culture medium for separating fungi and yeast-like fungi and preparation method thereof | |
| Hewlett | A Manual of bacteriology | |
| CN110343626A (en) | One plant of heat-resisting thermophilic Halomonas and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190726 |