CN104206416B - The application of hydrogen sulfide or its soluble-salt in collaboration enhancing sterilization effect - Google Patents
The application of hydrogen sulfide or its soluble-salt in collaboration enhancing sterilization effect Download PDFInfo
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- CN104206416B CN104206416B CN201410387400.2A CN201410387400A CN104206416B CN 104206416 B CN104206416 B CN 104206416B CN 201410387400 A CN201410387400 A CN 201410387400A CN 104206416 B CN104206416 B CN 104206416B
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- 229910000037 hydrogen sulfide Inorganic materials 0.000 title claims abstract description 80
- 238000004659 sterilization and disinfection Methods 0.000 title claims abstract description 49
- 230000001954 sterilising effect Effects 0.000 title claims abstract description 36
- 230000000694 effects Effects 0.000 title abstract description 36
- 230000002708 enhancing effect Effects 0.000 title abstract description 6
- 241000894006 Bacteria Species 0.000 claims abstract description 15
- 239000007800 oxidant agent Substances 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 9
- MHAJPDPJQMAIIY-UHFFFAOYSA-N hydrogen peroxide Substances OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 79
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- 238000007654 immersion Methods 0.000 claims description 5
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- Agricultural Chemicals And Associated Chemicals (AREA)
- Apparatus For Disinfection Or Sterilisation (AREA)
Abstract
The invention discloses the application of hydrogen sulfide or its soluble-salt in collaboration enhancing sterilization effect, the present invention fumigates or soaked article or utensil to be sterilized by using hydrogen sulfide, inactivate article or the anti-oxidant fermentoid of tool face bacterium, avoid decomposition of the anti-oxidant fermentoid to oxidant, bacterial death is made by the oxidative stress of oxidant again, the method substantially increases the death rate of microbial cell, enhances the effect of sterilization, shortens the time of sterilization.
Description
Technical field
Strengthen sterilization effect in collaboration the present invention relates to a kind of disinfectant, more particularly to hydrogen sulfide or its soluble-salt
In application.
Background technology
Sterilization and sterilizing are one of technologies the most frequently used during microorganism works.Medicine equipment, medicine, the production of food and
In application process, often experimental situation, relative article or whole workshop are carried out disinfection and sterilized.The method of sterilization has many
Kind, quite a few is to promote microbial cell dead based on oxidative stress.Such as hydrogen peroxide, potassium permanganate oxidant are killed
The sterilization of the halogen compounds such as bacterium, chlorine and calcium hypochlorite (bleaching powder), in addition ultraviolet disinfection and gamma radiation sterilization also with oxygen
Changing stress makes microbial cell death relevant.
Why oxidative stress can make microbial cell dead, be due to that can produce active oxygen (ROS) under oxidative stress.It is living
Property oxygen include various ingredients, wherein reactivity highest is hydroxy radical (HO), due to there is many iron content in bacterial cell
Albumen, these albumen can discharge Fe after by oxidative attacks2+, then with H2O2Reaction generation HO.Although the HO life-spans are very
It is short, but its chemical reactivity is high, can be with intracellular nearly all macromolecular substances (such as protein, DNA, RNA and fat
Matter) reacted, these large biological molecules are aoxidized, the damage and death of cell is caused.
Come from all kinds of active oxygens that extraneous or own metabolism is produced to resist, bacterium has evolved a variety of reply ways
Footpath.Most important of which is that cell can produce a series of energy decomposing Hs2O2Catalase.Intracellular H2O2Removing mainly by
Catalase and peroxidase are completed.In many bacteriums such as Escherichia coli (E.coli), AhpCF is main peroxide
Change hydrogen scavenger enzyme.AhpC is to H2O2With very high affinity, even if producing H under aerobic conditions2O2Speed be up to 15 μM/s,
AhpCF can also be by most of H2O2Decompose, make intracellular H2O2Stabilization is within 20nM.If but when the ring residing for bacterium
Border H2O2When concentration is more than 20 μM, due to H2O2Cell membrane can be readily passed through, intracellular H is caused2O2Amount has exceeded holding for AhpC
By scope, the cysteine residues in AhpC are made to be oxidized to sulfinic acid and inactivate.In addition, AhpCF removes H2O2Need consumption a large amount of
NADH, the energy charge of cell can be made excessive.Therefore, Cellular evolution goes out an other catalase-like.
Work as H2O2Amount beyond certain level just make cell be in oxidation state, this oxidation state can be by half Guang of some albumen
Histidine residue is perceived, and the H of nanomole level can be sensed if the oxidative stress response protein OxyREC in E.coli2O2。E.coli
In regulated and controled by OxyREC mainly have catalase katG, alkyl hydroperoxide reductases to the related gene of oxidation response
AhpCF, glutaredoxin grxA, glutathione reductase gorA, the aporepressor fur and DNA Protecting gene dps of iron balance
With RNA controlling genes oxyS.KatG is by H2O2Expressed topmost H after induction2O2Scavenger enzyme, it is a kind of difunctional
Enzyme, the activity of existing catalase has the activity of peroxidase again.KatG and H2O2With reference to Km values it is higher, even milli
The H of mole rank2O2Also it will not be caused to reach saturation；Need unlike AhpC other to go back the auxiliary of original system additionally, due to it
Help, thus intracellular NADH shortage will not be limited by.Due to intracellular AhpCF and KatG presence so that microorganism is external
The oxidative stress status on boundary has stronger resistivity.Sometimes it can not be still killed under higher oxidative pressure.So only
Carry out disinfection to sterilize by single oxidative bactericide and can not improve bactericidal effect, find a kind of anti-microbial agents of combination
The bactericidal effect of oxidative bactericide is improved, by as the effective means for improving bactericide bactericidal effect.
Publication No. discloses a kind of quick, efficient and environmentally friendly sterilizing side of desinfection chamber for CN102698309A patent
Method, this method is that the space that will be sterilized is cleaned out, right after the ClO 2 solution configured was activated through 20~30 minutes
The space of work carries out spraying treatment, then allows droplet to land naturally from space eminence, Seal treatment space immediately after spraying treatment
1h to 3h, is normally used subsequently into space.
The content of the invention
Present invention discover that hydrogen sulfide can suppress the vigor of catalase, bacterium life can be suppressed by being cooperateed with hydrogen peroxide
The formation of thing film, has obvious inhibiting effect to bacterial growth.
Found based on more than, the invention provides hydrogen sulfide or its soluble-salt in collaboration enhancing sterilization effect
Using.
The soluble-salt refers to water miscible sulfide or sulfohydrate, and the sulfide or sulfohydrate are being dissolved in water
After can generate hydrogen sulfide.
The enhancing sterilization effect refers to hydrogen sulfide or its soluble-salt and other disinfection way collective effects in treating
Sterilizing objects or utensil, are remained with the bacterium for reducing article or tool face.Hydrogen sulfide or its soluble-salt and other sterilization sides
Formula, can apply simultaneously, can also be first after-applied, preferably first handled with hydrogen sulfide or its soluble-salt, it is then used again
It carries out disinfection.
Present invention also offers a kind of sterilization method, with the article that sulfiding gas are stifling or hydrogen sulfide solution immersion is to be sterilized
Or utensil carries out disinfection again after for a period of time.
The sterilization refers to using disinfectant sterilization or radiosterilization.
The disinfectant can be oxidized form disinfectant, such as hydrogen peroxide, ozone, chlorine dioxide, Peracetic acid, bleaching
Powder, preferably hydrogen peroxide.
The radiosterilization can be disinfection by ultraviolet light, microwave sterilization or radiation sterilization.
The disinfectant sterilization or radiosterilization are to use existing conventional method.
Described stifling or immersion time is no less than 1 minute.
Final concentration of 0.1~the 4g/m of hydrogen sulfide that the hydrogen sulfide gas is fumigated or hydrogen sulfide solution soaks3。
The present invention makes article or tool face bacterium by the way that hydrogen sulfide is stifling or soak article or utensil to be sterilized
Anti-oxidant fermentoid inactivation, it is to avoid decomposition of the anti-oxidant fermentoid to oxidant, then bacterium is made by the oxidative stress of oxidant
Death, the method substantially increases the death rate of microbial cell, enhances the effect of sterilization, also shortens sterilization
Time.
Brief description of the drawings
Fig. 1 combines bent to the growth inhibition of Ao Naide Shewanellas with hydrogen peroxide for the various concentrations hydrogen sulfide of embodiment 1
Line.
1：Compare (plus equivalent sterilized water)；2：0.1mM H2S；3：0.25mM H2O2；4：12.5μM H2S+0.25mM
H2O2；5：25μM H2S+0.25mM H2O2；6：50μM H2S+0.25mM H2O2。
Fig. 2 embodiments 2 combine the effect to pseudomonas aeruginosa Synergistic biocidal for various concentrations hydrogen sulfide with hydrogen peroxide
Figure.
1：0μM Na2S+0.25mM H2O2；2：10μM Na2S+0.25mM H2O2；3：20μM Na2S+0.25mM H2O2；
4：50μM Na2S+0.25mM H2O2；5：100μM Na2S+0.25mM H2O2。
Fig. 3 is that the various concentrations hydrogen sulfide of embodiment 3 is combined with different hydrogen peroxide to Ao Naide Shewanella Synergistic biocidals
Design sketch.
Pitch black post is from left to right：0.1mM H2S+0；0.125mM；0.25mM；0.5mM and 1mM H2O2。
Somber post is from left to right：0.；0125mM；0.025mM；0.05mM and 0.1mM H2S+0.25mM H2O2。
Fig. 4 is that the hydrogen sulfide of embodiment 4 cooperates with influence curve with hydrogen peroxide to bacillus subtilis bacteria growing, (A) shaking flask
Culture；(B) static gas wave refrigerator.
Fig. 5 is that the hydrogen sulfide of embodiment 5 cooperates with influence curve with hydrogen peroxide to Escherichia coli Growth, (A) Shaking culture；
(B) static gas wave refrigerator.
Fig. 6 is influence post of the various concentrations hydrogen sulfide of embodiment 6 to Ao Naide Shewanella catalase relative activities
Shape figure.
Fig. 7 is influence curve of the various concentrations hydrogen sulfide of embodiment 6 to calf liver catalase relative activity.
Fig. 8 is 24 orifice plates addition Na in embodiment 72S and H2O2Suppress the photo of biofilm formation afterwards.
Fig. 9 is that the collaboration influence that the various concentrations hydrogen sulfide of embodiment 7 is combined with hydrogen peroxide on bacterial biof iotalm formation is bent
Line.
Embodiment
The coordinate repression that the hydrogen sulfide of embodiment 1 grows with hydrogen peroxide to Ao Naide Shewanellas
Ao Naide Shewanellas (S.oneidensis) LB culture mediums (yeast extract 0.5%, peptone 1%,
NaCl0.5%, pH7.2) in 30 DEG C of 200rpm overnight incubations, by 0.1mL cultures access 3.7mLLB culture mediums in, then add
0.1mL NaHS solution and 0.1mLH2O2(control is replaced with water), it is 4mL to make cumulative volume.Experiment be arranged in parallel 6 groups, every group 3
Repeat.Except one group of blank control (plus 0.2mL sterilized waters) and 2 groups of single controls (are respectively 0.1mM H2S and 0.25mM H2O2)
Outside, 3 groups of cooperative experiment H2S and H2O2Final concentration be respectively：12.5μM H2S+0.25mM H2O2、25μM H2S+0.25mM
H2O2With 50 μM of H2S+0.25mM H2O2.30 DEG C are unified in after sample-adding, is cultivated under 200rpm, culture is determined per hour and is existed
600nm absorbance (OD600), as a result (average value) is as shown in Figure 1.
As can be seen that individually adding H2S and H2O2Fungistatic effect is very poor, but H2S and H2O2There is fungistatic effect simultaneously obvious
Enhancing, and with H2The raising of S concentration, fungistatic effect also strengthens therewith, works as H2When S concentration increases to 50 μM, inhibit substantially
The growth of Ao Naide Shewanellas.
The hydrogen sulfide of embodiment 2 and synergic remove of the hydrogen peroxide to pseudomonas aeruginosa
37 DEG C of 200rpm of pseudomonas aeruginosa are cultivated to OD600=0.2,100 μ L are inhaled, 880 μ L sterile salines are added to
In, then add 10 μ L Na2S solution and 10 μ L H2O2, it is 1mL to make cumulative volume, and experiment be arranged in parallel 5 groups, every group of 3 repetitions.5 groups
The hydrogen sulfide final concentration of experiment is respectively 0,0.0125mM, 0.025mM, 0.05mM and 0.1mM, adds Na2S solution simultaneously shakes up
Afterwards, H is added at once2O2, and make its final concentration all be 0.25mM.After shaking up, handled 30 minutes in 30 DEG C of insulating boxs, cooled on ice,
Gradient dilution is carried out at once, and by 10-2, 10-3, 10-4, 10-5, 10-6Each 5 μ L points of dilution on LB flat boards, in 30 DEG C of perseverances
Cultivated 20 hours in incubator, concrete outcome is as shown in Figure 2.
As can be seen that H is used alone2O2, bactericidal effect is not good enough, but with H2The raising of S concentration, hydrogen sulfide and peroxidating
Hydrogen is more and more stronger to the Synergistic biocidal effect of pseudomonas aeruginosa, works as H2S concentration reaches 100 μM, and verdigris has almost been killed completely
Pseudomonad.
The hydrogen sulfide of embodiment 3 and synergic remove of the hydrogen peroxide to Ao Naide Shewanellas
30 DEG C of 200rpm of Ao Naide Shewanellas are cultivated to OD600=0.35,100 μ L are inhaled, 880 μ L sterile physiological salt are added to
In water, then add 10 μ L NaHS solution and 10 μ L H2O2, it is 1mL to make cumulative volume, and experiment be arranged in parallel 10 groups, every group of 3 repetitions.
Wherein 5 groups of hydrogen sulfide final concentration is respectively 0,0.0125mM, 0.025mM, 0.05mM and 0.1mM, H2O2Final concentration be all
0.25mM.Other 5 groups of concentration of hydrogen sulfide is fixed as 0.1mM (final concentration), H2O2Final concentration be respectively set to 0,0.125mM,
0.25mM, 0.5mM and 1mM, after shaking up, handle 30 minutes, cooled on ice in 30 DEG C of insulating boxs, carry out gradient dilution simultaneously at once
Inhale 10-4, 10-5, 10-6Each 0.1mL of dilution be coated on LB flat boards, in 30 DEG C of insulating boxs cultivate 24 hours after count single bacterium
Fall, and calculate the viability of bacterium, concrete outcome is as shown in Figure 3.
As can be seen that H is used alone2O2, bactericidal effect is good, but with H2The raising of S concentration, hydrogen sulfide and peroxidating
Hydrogen is more and more stronger to the bactericidal effect of Ao Naide Shewanellas.
The growth inhibition effect of the hydrogen sulfide of embodiment 4 and hydrogen peroxide to bacillus subtilis
Bacillus subtilis (Bacillus subtilis) 37 DEG C of 200rpm overnight incubations in LB culture mediums, 0.1mL trainings
Foster thing is seeded in 3.7mL LB culture mediums, then adds 10 μ L NaHS solution and 10 μ L H2O2, it is 4mL to make cumulative volume, is tested parallel
4 groups, every group of 3 repetitions are set.Blank control adds 20 μ L H2O, single-factor control respectively adds 10 μ L H2H in O, experiment2S and H2O2's
Final concentration is respectively：0.05mM and 1mM.Static at 37 DEG C or shaking flask (200rpm) is cultivated after mixing, and growth is determined per hour
Measure (OD600), concrete outcome (average value) is as shown in Figure 4.
As shown in Figure 4, bacillus subtilis is less sensitive to hydrogen peroxide, and more sensitive to hydrogen sulfide, to this kind of antioxygen
The stronger bacterial strain of change ability, first with hydrogen sulfide treatment, can increase its sensitiveness to oxidant.
The growth inhibition effect of the hydrogen sulfide of embodiment 5 and hydrogen peroxide to Escherichia coli
Escherichia coli (E.coli) 37 DEG C of 200rpm overnight incubations in LB culture mediums, 0.1mL cultures are seeded to 3.7mL
In LB, then add 10 μ L NaHS solution and 10 μ L H2O2, it is 4mL to make cumulative volume, and experiment be arranged in parallel 4 groups, every group of 3 repetitions.It is empty
White control plus 20 μ L H2O, single-factor control respectively adds 10 μ L H2H in O, experiment2S and H2O2Final concentration be respectively：0.05mM and
1mM.Static at 37 DEG C or shaking flask (200rpm) is cultivated after mixing, and increment (OD is determined per hour600), concrete outcome is (average
Value) as shown in Figure 5.
As shown in Figure 5, H is used alone2S or H2O2It is poor to the fungistatic effect of Escherichia coli, illustrate Escherichia coli to H2S
And H2O2It is all insensitive, the fungistatic effect of Escherichia coli is remarkably reinforced when both are used in mixed way, fungistatic effect is more during static gas wave refrigerator
Substantially.
The hydrogen sulfide of embodiment 6 suppresses catalase activity
Ao Naide Shewanella 200rpm 30C Shaking cultures 4 hours are to OD600=0.45, experiment be arranged in parallel 6 groups, often
3 repetitions of group.4mL nutrient solutions are inhaled, after phosphate buffer is from washing, plus 1mL phosphate buffers, sonicated cells (work 3 seconds,
Gap 4 seconds, 20 circulations), after centrifugation, the μ L of supernatant (crude enzyme liquid) 180 are taken, 10 μ L NaHS solution and 10 μ L H are added2O2, make total
Volume is 200 μ L, H therein2S concentration is respectively 0 μM, 12.5 μM, 25 μM, 50 μM, 100 μM and 200 μM, H2O2For 500 μM.
After 30 DEG C of insulation 30min, centrifugation, plus 1500 μ L FOX-1 reagents, reacted at room temperature 30 minutes after mixing, survey OD560(H2O2With FOX-
1 reagent reacting, H2O2It is more, OD560It is higher), seek remaining hydrogen peroxide.According to the H of consumption2O2Amount, calculates catalase
Relative activity, as a result see Fig. 6.
It will be appreciated from fig. 6 that hydrogen sulfide can significantly inhibit the vigor of bacterium hydrogen peroxide enzyme, so as to reduce catalase
Degraded to hydrogen peroxide, reduces the reduction of concentration of hydrogen peroxide in solution.
The cell extract of bacterium, remaining behaviour are replaced with small beef liver catalase standard items (being purchased from bio-engineering corporation)
Make ibid, detection (average value) result is as shown in Figure 7.
As shown in Figure 7, the H in initial soln2O2For 500 μM, after being reacted through certain time, remaining peroxidating in solution
Hydrogen (A560) be directly proportional to concentration of hydrogen sulfide, illustrate that hydrogen sulfide significantly inhibits small beef liver catalase, 50 μM of hydrogen sulfide is almost
Completely inhibit small beef liver catalase vigor.
The hydrogen sulfide of embodiment 7 cooperates with the formation for suppressing bacterial biof iotalm with hydrogen peroxide
2ml LB culture mediums are added in 24 orifice plates, and Na is added as shown in Figure 8 in every hole2S and H2O2Solution, makes the H in every hole2S
And H2O2Final concentration reach setting value, exposure 30 minutes in outdoor elements, 30 DEG C of cultures are taken pictures after 24 hours, and are received
Obtain and mixed after the biomembrane to be formed, plus sterilized water 2mL, determine the biomass (OD of film600), as a result (average value) is shown in Fig. 9.
From Fig. 8 and Fig. 9, without H2O2In the case of, H2The no inhibitory action of formation of the S to biomembrane, and add
Plus H2O2Afterwards, with H2The raising of S concentration, the generation to bacterial biof iotalm, inhibitory action is stronger.Equally, without H2S's
In the case of, H2O2To the no inhibitory action of formation of biomembrane, and add H2After S, with H2The raising of S concentration, to bacterium living beings
The inhibitory action of film is stronger.
The hydrogen sulfide of embodiment 8 cooperates with the Disinfection Effect handled to desinfection chamber with hydrogen peroxide
It is 50m in volume3Desinfection chamber in, first using 4g/m3Hydrogen sulfide is handled desinfection chamber and maintained 10 minutes,
10g/m is used again3Hydrogen peroxide steam is handled and maintained 60 minutes, filtrated air is passed through after terminating 10 minutes, sterile LB is put down
Plate after 30 minutes, is cultivated 24 hours exposed to desinfection chamber in 30 DEG C of insulating box, the clump count in detection LB flat boards, only to adopt
Use 10g/m3Hydrogen peroxide steam maintains the disinfection way of 60 minutes to compare, and evaluates the sterilization effect of two kinds of processing, often
Kind of processing be arranged in parallel three groups of experiments, averaging, as a result as shown in table 1.
The hydrogen sulfide of table 1. cooperates with the Disinfection Effect handled to desinfection chamber with hydrogen peroxide
As shown in Table 1, the sterilization effect of desinfection chamber is substantially better than under the synergy of hydrogen sulfide and hydrogen peroxide
Only use the effect of disinfectant with hydrogen peroxide.
The hydrogen sulfide of embodiment 9 cooperates with the Disinfection Effect handled to ultra-clean chamber article with gaseous oxidizer
It is about 1.2m in volume3, temperature is in 25 DEG C of ultra-clean chamber, stacks the culture dish of sterilizing to be sterilized, test tube and rubber
Sebific duct, first using 0.1g/m3Hydrogen sulfide is handled ultra-clean chamber and maintained 10 minutes, then using 5g/m3Gaseous oxidizer processing
And maintain 30 minutes, filtrated air is passed through after terminating 10 minutes, hydrogen peroxide steam, ozone is respectively adopted in above-mentioned gaseous oxidizer
Steam, Peracetic acid steam and chlorine dioxide steam, to be compared only with the disinfection way of gaseous oxidizer.After end, use
50mL sterilized waters rinse these sterilizing articles 50 times repeatedly, then by washings be mixed into 450mL be incubated in 55 DEG C LB solids training
Support in base, plate is down flat after mixing, per ware 25mL, after flat board is cultivated 24 hours in 30 DEG C of insulating box, 20 LB of detection are put down
Total clump count in plate, to evaluate sterilization effect, every kind of processing be arranged in parallel three groups of experiments, averaging, as a result such as table 2
It is shown.
The different disinfectants of table 2. are cooperateed with hydrogen sulfide to be compared the Disinfection Effect of ultra-clean chamber article
As shown in Table 2, the bactericidal effect that ultra-clean chamber is sterilized under hydrogen sulfide and gaseous oxidizer combination is better than alone gaseous state
The sterilization effect of oxidant sterilization.
The hydrogen sulfide of embodiment 10 cooperates with the Disinfection Effect handled to superclean bench with ultraviolet
Superclean bench：About 1 cubic metre of volume, is sprayed with ultraviolet irradiation 30 minutes and 10mL0.01mM hydrogen sulfide respectively
Afterwards, then with ultraviolet irradiation 30 minutes handle, ibid with plate exposure method detection sterilization effect, every kind of processing be arranged three in parallel
Group experiment, averaging the results are shown in Table 3.
The hydrogen sulfide of table 3. cooperates with the Disinfection Effect handled to superclean bench with hydrogen peroxide
| Processing | Clump count 1 | Clump count 1 | Clump count 1 | It is average |
| Ultraviolet irradiation 30 minutes | 20 | 28 | 16 | 21 |
| 0.01mM hydrogen sulfide+ultraviolet irradiation 30 minutes | 0 | 2 | 1 | 1 |
As shown in Table 3, hydrogen sulfide can also strengthen the sterilization effect of ultraviolet.
The hydrogen sulfide of embodiment 11 cooperates with the Disinfection Effect handled to tableware with bleaching powder
5% bleaching powder of lunch box soaks immersion in processing in 20 minutes and 100 μM of hydrogen sulfide solutions and floated again with 5% after 5 minutes
White powder, which soaks 15 minutes, to be handled, and is rinsed twice with 100mL sterile salines after processing, plus the concussion of 10mL sterile salines,
Collection swings the physiological saline after washing and it is enriched in into miillpore filter, is affixed on LB flat boards, sterilization effect is detected after culture,
Every kind of to handle the three groups of experiments that be arranged in parallel, averaging the results are shown in Table 4.
The hydrogen sulfide of table 4. cooperates with the Disinfection Effect handled to tableware with bleaching powder
| Processing | Clump count 1 | Clump count 1 | Clump count 1 | It is average |
| Bleaching powder | 48 | 32 | 35 | 38 |
| The 100 μM of bleaching powder of hydrogen sulfide+5% immersions | 7 | 5 | 8 | 7 |
As shown in Table 4, hydrogen sulfide can strengthen the sterilization effect of bleaching powder.
Claims (1)
1. a kind of method for disinfection and sterilization, it is characterised in that in volume be 50m3Desinfection chamber in, first using 4g/m3Hydrogen sulfide is to nothing
Bacterium room is handled and maintained 10 minutes, then using 10g/m3Hydrogen peroxide steam is handled and maintained 60 minutes, is passed through after terminating
Filtrated air 10 minutes；
Or, it is 1.2m in volume3, temperature in 25 DEG C of ultra-clean chamber, first using 0.1g/m3Hydrogen sulfide to ultra-clean chamber at
Manage and maintain 10 minutes, then using 5g/m3Gaseous oxidizer is handled and maintained 30 minutes, and 10 points of filtrated air is passed through after terminating
Clock；
The gaseous oxidizer is hydrogen peroxide steam, ozone steam, Peracetic acid steam or chlorine dioxide steam；
Or, in the superclean bench of 1 cubic metre of volume, after being sprayed with 10mL 0.01mM hydrogen sulfide, then with ultraviolet irradiation 30
Minute；
Or, by lunch box with 100 μM of hydrogen sulfide solutions soak 5 minutes after, then with 5% bleaching powder immersion 15 minutes.
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