CN101363003B - Microorganism resistance test apparatus and method for verification of plasma sterilization - Google Patents

Microorganism resistance test apparatus and method for verification of plasma sterilization Download PDF

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CN101363003B
CN101363003B CN2008101210088A CN200810121008A CN101363003B CN 101363003 B CN101363003 B CN 101363003B CN 2008101210088 A CN2008101210088 A CN 2008101210088A CN 200810121008 A CN200810121008 A CN 200810121008A CN 101363003 B CN101363003 B CN 101363003B
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plasma
verification
tank
vacuum
resistance test
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CN101363003A (en
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刘川生
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Liu Chuansheng
Zhang Jian
Zhang Liubo
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Abstract

The invention relates to a microbial resistance test device and a method for plasma sterilization verification. The technical problem to be solved is to provide the microbial resistance test device and the method for the plasma sterilization verification, the invention aims at carrying out the research of the related sterilization resistance of standard microbial strains which are used after the hydrogen peroxide plasma sterilization test through the device and the method, thereby comparing and selecting the strain with the strongest resistance as the standard test strain. The technical proposal for solving the problems is as follows: a resistance detection device is arranged in an experimental tank room, a temperature sensor II is matched therewith, a neutralizer filling device and a sterilization media filling device are connected at the outside of the experimental tank room, the neutralizer filling device is communicated with the resistance detection device through a connecting guide tube I via a sealing interface on the wall of the experimental tank room, and the sterilization media filling device is connected with an atomizing nozzle which is arranged above the tank wall of avacuum tank through a connecting guide tube II via a micro-electromagnetic valve. The microbial resistance test device and the method can be used for the hydrogen peroxide plasma sterilization test.

Description

The microorganism resistance test apparatus and the method that are used for verification of plasma sterilization
Technical field
The present invention relates to the drag experimental installation, especially a kind of microorganism resistance test apparatus and method that is used for verification of plasma sterilization.Not only be applicable to the hydrogen peroxide plasma sterile test, also be applicable to the sterilization drag test of the microbial strains of various medical cryogenic gas sterilising method checking.
Background technology
The hydrogen peroxide plasma temperature sterilization is at present up-to-date in the world a kind of medical temperature sterilization technology; With other conventional medical sterilization compared with techniques; Have characteristics such as low temperature is quick, energy-conserving and environment-protective, economic security, can be used for clinical departments such as hospital operating room, Supply House, sight glass chamber various medical devices (comprising metallic weapon and nonmetal apparatus) is carried out quick temperature sterilization processing.
As medical sterilizing device, the checking that need make an experiment to the sterilization usefulness of this equipment and technological method, this just need carry out proof test through the selected the strongest standard bacterial classification of a kind of drag of correlation test.And selected which kind of bacterial classification; Then need measure multiple microbial strains the drag index D value of this sterilising method (is killed the needed time of 90% mikrobe; The D value is big more; Explain that this bacterial classification is strong more to the drag of this sterilising method), thus relatively and select the strongest bacterial classification of drag as the standard test bacterial classification, the usefulness proof test is used to sterilize.Because hydrogen peroxide plasma temperature sterilization technology is a new technology, at present, still there is not a kind of easy, reliable testing apparatus to carry out the drag test of above-mentioned checking bacterial classification.
Summary of the invention
The technical problem that the present invention will solve is: a kind of microorganism resistance test apparatus and method that is used for verification of plasma sterilization is provided; Be intended to through these apparatus and method used standard microorganism bacterial classification carries out the research of the relevant drag of sterilizing through hydrogen peroxide plasma sterilization test back, thereby relatively and select the strongest bacterial classification of drag as the standard test bacterial classification.
The technical scheme that the present invention adopted is: the microorganism resistance test apparatus that is used for verification of plasma sterilization; Have vacuum tank and hermatic door; Thereby form the experimental tank chamber; Inwall in the experimental tank chamber is equipped with discharge electrode and is connected with coupling tuner and plasma exciatiaon power supply with concentric cable through the sealing feed-in part on the tank body, and the plasma exciatiaon power supply is connected with electric apparatus control apparatus again; The sidewall of said vacuum tank is provided with and vacuumizes interface; This interface is connected with vacuum pump through the high vacuum flapper valve; The other end connecting circuit gear of vacuum pump; Also be furnished with the inflation interface on the said tank skin, this interface is connected the other end connecting circuit gear of high vacuum flapper valve through the high vacuum flapper valve with air filter; Also equipped electrothermic constant temperature equipment and vacuum detecting transmitter on the outer side wall of said vacuum tank; It is characterized in that said experimental tank is indoor is equipped with the drag proofing unit and is furnished with TP II; Be connected to neutralizing agent charging point and sterile medium charging point in the outside of experimental tank chamber; Said neutralizing agent liquid-adding device is communicated with the drag proofing unit through the sealed interface of linking conduit I on the experimental tank locular wall, and said sterile medium filling apparatus is connected to the dispersive nozzle that is located at vacuum tank tank skin top through linking conduit II through miniature electromagnetic valve.
Said drag proofing unit has the mechanical drive that is fixed on middle part, experimental tank chamber; This device is made up of direct-current machine, wheel box, turn-screw, driving nut, portable bar, axle sleeve and overcoat; Outer putting in its vertical fixing is fixed with lower fixed frame and at the one of which end movable block down is housed; Be fixed with upper holder on the portable bar of mechanical drive and movable block and micro valve be housed at the one of which end; Portable bar outside on upper holder is with spring and on its top gland is housed, and between this gland and micro valve, is provided with fixed briquetting and active spill spin block.
Said electrothermic constant temperature equipment is made up of the hot plate and the thermal insulation layer that are located at the vacuum tank outside, and is furnished with TP and temperature controller.
Said vacuum tank is processed by stainless steel, titanium alloy or duraluminum, is shaped as drum shape or rectangle, vertical or horizontal placement, and volume is 30~150L.
Said discharge electrode is processed by stainless steel, titanium alloy or duraluminum porous plate, with the tank skin gap be 5~50mm, and insulate with tank skin.
Said neutralizing agent charging point comprises 1~5 syringe and liquid feeding carriage, and each syringe has the substratum that contains neutralizing agent of 5~10ml.
Said sterile medium charging point comprises syringe and liquid feeding carriage.
Near said neutralizing agent charging point and the sterile medium charging point TP I is housed.
A kind of microorganism resistance TP that is used for verification of plasma sterilization is characterized in that comprising the steps:
The substratum 10mL that 1) will contain neutralizing agent is one by one in 1~5 syringe of suction; Be installed in respectively on the liquid feeding carriage; Zylox linking conduit I with internal diameter 2mm is connected the rubber seal plug on sealed interface on syringe, the vacuum tank and the withstand voltage test tube; With 1~5 test organisms sheet be put into respectively 1~5 in vitro withstand voltage, cover rubber seal plug;
The withstand voltage test tube that 2) the test organisms sheet will be housed is installed on the lower fixed frame of drag proofing unit, clamps with movable block down, and rubber seal plug is used movable block and upper holder is fixed, and said Zylox linking conduit I is connected with micro valve;
3) with disinfectant H 2O 2In the syringe of suction sterile medium charging point, be installed on the liquid feeding carriage, use internal diameter to be connected on the miniature electromagnetic valve as the tetrafluoroethylene linking conduit II of 1.5mm;
4) close the hermatic door of experimental tank chamber, on testing apparatus control touch-screen, set 0~999 second grouping time, each bacterial classification is divided into 10 groups according to germicidal action time length and makes an experiment, every group of 1~5 sample;
5) the open vacuum pump vacuumizes, and opens the rubber seal plug on the withstand voltage test tube;
6) when vacuum tightness reaches 50~150Pa, close the sealing plug of withstand voltage test tube, open the plasma exciatiaon power supply and carried out preparatory plasma glow discharge 1~10 minute;
7) open miniature electromagnetic valve, add disinfectant H 2O 2And vaporization, keep diffusion in 1~10 minute;
8) sealing plug of opening withstand voltage test tube carries out bactericidal reaction and timing, and the reaction times was controlled at 0~999 second, immediately closed the sealing plug of withstand voltage test tube, opened micro valve simultaneously, made neutralizing agent submergence bacterium sheet, and the time control accuracy was 0.1 second, stopped reaction;
9) be evacuated down to 50~150Pa once more, open the plasma exciatiaon power supply, plasma discharge 1~10 minute;
10) close the plasma exciatiaon power supply, open the inflation interface, the inflation concora crush;
11) open hermatic door, take out the withstand voltage test tube of band sealing plug;
12) sterilization finishes, and gets 1~3 at random 1~5 bacterium sheet the inside and carries out the enumeration of bacterium sheet.
The invention has the beneficial effects as follows: the present invention provides a kind of microorganism resistance test apparatus and method for the technical identification of hydrogen peroxide plasma temperature sterilization; Can optimize the strongest bacterial classification of drag easy, reliably through this testing apparatus, thereby carry out the experimental study of relevant sterilising effect checking as the standard test bacterial classification.
Description of drawings
Fig. 1 is that structure of the present invention is always schemed.
Fig. 2 is the structure iron (rubber seal plug is in sealed state) of a kind of state of drag proofing unit among the present invention.
Fig. 3 is the structure iron (rubber seal plug is in opened condition) of the another kind of state of drag proofing unit among the present invention.
Embodiment
Like Fig. 1-shown in Figure 3; Present embodiment has vacuum tank 1 and hermatic door 2; Thereby form experimental tank chamber 3; Inwall in the experimental tank chamber is equipped with discharge electrode 4 and is connected with coupling tuner 9 and plasma exciatiaon power supply 10 with concentric cable through the sealing feed-in part on the tank body, and the plasma exciatiaon power supply is connected with electric apparatus control apparatus 12 again.Said discharge electrode 4 is processed by metallic porous sheets such as stainless steel, titanium alloy or duraluminums; With the tank skin gap be 5~50mm; And with tank skin insulation, so-called plasma exciatiaon discharge is exactly to pass through discharge electrode 4, under the control of program in the photoglow of the indoor formation plasma body of test tank.Output power of power supply is adjustable in 50~500W scope, and discharge time, control accuracy was ± 0.1 second.
The sidewall of said vacuum tank 1 is provided with and vacuumizes interface 6, and this interface is connected the other end connecting circuit gear 12 of vacuum pump through high vacuum flapper valve 8 with vacuum pump 7; Also be furnished with inflation interface 24 on the said tank skin, this interface is connected the other end connecting circuit gear 12 of high vacuum flapper valve 8 through high vacuum flapper valve 8 with air filter 11.Vacuum control unit can be made to vacuumize in the jar to reach specified vacuum value (30~300Pa) by time variable control open vacuum pump and each valve fast.Being provided with of inflation interface 24 can make testing laboratory recover concora crush in a short period of time, so that opening encapsulation door and take out withstand voltage test tube 21 fast.
Also equipped electrothermic constant temperature equipment 5 and vacuum detecting transmitter 13 on the outer side wall of said vacuum tank 1; This electrothermic constant temperature equipment 5 is made up of the hot plate and the thermal insulation layer that are located at vacuum tank 1 outside; And be furnished with TP and temperature controller, the warranty test chamber wall temperature is controlled in ℃ scope of room temperature~80.The temperature display precision is 0.1 ℃.
Drag proofing unit 20 is installed in the said experimental tank chamber 3 and is furnished with TP II14-2.Said drag proofing unit 20 has the mechanical drive 23 that is fixed on 3 middle parts, experimental tank chamber; This device is made up of direct-current machine 23-1, wheel box 23-2, turn-screw 23-3, driving nut 23-4, portable bar 23-5, axle sleeve 23-6 and overcoat 23-7; On the overcoat 23-7 of its vertical fixing, be fixed with lower fixed frame 20-9 and movable block 20-1 down be housed at the one of which end; On the portable bar 23-5 of mechanical drive 23, be fixed with upper holder 20-8 and movable block 20-2 and micro valve 20-3 be housed at the one of which end; Portable bar 23-5 outside on upper holder 20-8 is with spring 20-7 and on its top gland 20-6 is housed, and between this gland and micro valve 20-3, is provided with fixed briquetting 20-4 and active spill spin block 20-5.
In order accurately to finish sterilization process; Eliminate the subsequent affect of sterile medium to mikrobe; Need set according to test and require at regular time and quantity at the withstand voltage neutralizing agent of in vitro annotating; Therefore, this example is connected to neutralizing agent charging point 16 in the outside of experimental tank chamber 3, and said neutralizing agent liquid-adding device is communicated with the drag proofing unit through the sealed interface of linking conduit I15-1 on the experimental tank locular wall.Said neutralizing agent charging point 16 comprises 5 syringes and liquid feeding carriage, and each syringe has the substratum that contains neutralizing agent of 5~10ml.When the drag test stopped, program was opened the micro valve 20-3 on the withstand voltage test tube sealing plug automatically, neutralizing agent is joined in vitro, and submergence bacterium sheet was to reach the purpose that stops bactericidal reaction.
Sterile medium of the present invention is a hydrogen peroxide liquid, becomes gaseous state again and evenly spreads after need in sterilization process, joining the indoor and atomizing of experimental tank at regular time and quantity, realizes the purpose of killing microorganisms.Therefore, be connected to sterile medium charging point 17 in the outside of experimental tank chamber 3, this sterile medium charging point 17 is connected to the dispersive nozzle 19 that is located at vacuum tank tank skin top through linking conduit II15-2 through miniature electromagnetic valve 18.Said sterile medium charging point 17 comprises syringe and liquid feeding carriage.Time requirement according to test is set is controlled automatic liquid feeding by miniature electromagnetic valve, and time control can be accurate to 0.1 second, and this sterile medium charging point can be controlled at accurate and adjustable in 0.1~5ml scope with the hydrogen peroxide IR.
Said vacuum tank 1 is processed by metallic substance such as stainless steel, titanium alloy or duraluminums, is shaped as drum shape or rectangle, vertical or horizontal placement, and volume is 30~150L; Be furnished with metal or nonmetal vacuum-sealing door 2, it is convenient to open;
Near said neutralizing agent charging point 16 and the sterile medium charging point 17 TP I14-1 is housed.
Electric apparatus control apparatus 12 in this example constitutes control circuit board by micro-chip unicircuit and color liquid crystal touch display screen; Constitute automatic running gear through functional devices such as rly. and alternating current contactors again; Through the program software in the singlechip chip, realize the automatic operating of complete assembly system.Said program software is the basic demand according to the test of microbiology sterilising plant drag, in conjunction with the characteristics and the function of apparatus of the present invention, and the specific routine software of being worked out, and pass through the automatic operation that single-chip computer control system is realized complete assembly.
Present embodiment is a kind of microorganism resistance TP that is used for verification of plasma sterilization, has comprised following steps:
The substratum 10mL that 1) will contain neutralizing agent is one by one in 5 (5 of this examples are one group) syringes of suction; Be installed in respectively on the liquid feeding carriage; With the Zylox linking conduit I15-1 of internal diameter 2mm the interface of sealed interface on syringe, the vacuum tank 1 and the rubber seal plug 22 on the withstand voltage test tube 21 is connected; 5 test organisms sheets are put into respectively in 5 withstand voltage test tubes 21, cover rubber seal plug 22.
Requirement according to " disinfection technology standard " is processed the bacterium sheet with test strain gemma suspension.Require the bacterium amount 1 * 10 6~5 * 10 6The CFU/ sheet need carry out enumeration.
The withstand voltage test tube 21 that 2) the test organisms sheet will be housed is installed on the lower fixed frame 20-9 of drag proofing unit 20; Clamp with following movable block 20-1; Rubber seal plug 22 is used movable block 20-2 and upper holder 20-8 fixes, said Zylox linking conduit I15-1 is connected with micro valve 20-3.
3) with disinfectant H 2O 2In the syringe of suction sterile medium charging point 17, be installed on the liquid feeding carriage, use internal diameter to be connected on the miniature electromagnetic valve 18 as the tetrafluoroethylene linking conduit II15-2 of 1.5mm.
4) close the hermatic door 2 of experimental tank chamber 3, on testing apparatus control touch-screen, set 0~999 second grouping time, each bacterial classification is divided into 10 groups according to germicidal action time length and makes an experiment, every group of 5 samples.
5) open vacuum pump 7 vacuumizes, and opens the rubber seal plug 22 on the withstand voltage test tube 21.
6) when vacuum tightness reaches 50~150Pa, close the sealing plug of withstand voltage test tube, open plasma exciatiaon power supply 10 and carried out preparatory plasma glow discharge 1~10 minute.
7) open miniature electromagnetic valve 18, add disinfectant H 2O 2And vaporization, keep diffusion in 1~10 minute.
8) sealing plug of opening withstand voltage test tube carries out bactericidal reaction and timing, and the reaction times was controlled at 0~999 second, immediately closed the sealing plug of withstand voltage test tube; Open micro valve 20-3 simultaneously; Make neutralizing agent submergence bacterium sheet, the time control accuracy is 0.1 second, stopped reaction.
9) be evacuated down to 50~150Pa once more, open plasma exciatiaon power supply 10, plasma discharge 1~10 minute.
10) close plasma exciatiaon power supply 10, open inflation interface 24, the inflation concora crush.
11) open hermatic door 2, take out the withstand voltage test tube of band sealing plug.
12) sterilization finishes, and gets 3 at random 5 bacterium sheet the insides and carries out the enumeration of bacterium sheet.
Enumeration is exactly to calculate the logarithmic value of organizing average survival gemma on the sample each action time.With action time is X-coordinate (X), and the logarithmic value of survival gemma is ordinate zou (Y), calculates gemma survival and the regression equation (Y=a+bX) of action time.Calculate the degree of correlation of each practical measurement value and linear regression equation,, calculate and reduce 90% gemma needed action time of D value according to the gained linear regression equation.
Set the different programs parameter, can carry out the drag test under multiple bacterial classification, the multiple condition.Changing other sterile medium composition of adding, can be liquid (like formaldehyde, Peracetic Acid etc.), also can be gas (oxyethane, oxygen, carbonic acid gas etc.), can carry out the sterilising effect simultaneous test after different media forms plasma body.

Claims (9)

1. microorganism resistance test apparatus that is used for verification of plasma sterilization; Have vacuum tank (1) and hermatic door (2); Thereby form experimental tank chamber (3); Inwall in the experimental tank chamber is equipped with discharge electrode (4) and is connected with coupling tuner (9) and plasma exciatiaon power supply (10) with concentric cable through the sealing feed-in part on the tank body, and the plasma exciatiaon power supply is connected with electric apparatus control apparatus (12) again; The sidewall of said vacuum tank (1) is provided with and vacuumizes interface (6); This interface is connected with vacuum pump (7) through high vacuum flapper valve (8); The other end connecting circuit gear (12) of vacuum pump; Also be furnished with inflation interface (24) on the said sidewall, this interface is connected the other end connecting circuit gear (12) of high vacuum flapper valve through high vacuum flapper valve (8) with air filter (11); Also equipped electrothermic constant temperature equipment (5) and vacuum detecting transmitter (13) on the outer side wall of said vacuum tank (1); It is characterized in that: drag proofing unit (20) is installed in the said experimental tank chamber (3) and is furnished with TP II (14-2); Be connected to neutralizing agent charging point (16) and sterile medium charging point (17) in the outside of experimental tank chamber (3); Said neutralizing agent charging point is communicated with the drag proofing unit through the sealed interface of linking conduit I (15-1) on the experimental tank locular wall, and said sterile medium charging point (17) is connected to the dispersive nozzle (19) that is located at vacuum tank tank skin top through linking conduit II (15-2) through miniature electromagnetic valve (18).
2. the microorganism resistance test apparatus that is used for verification of plasma sterilization according to claim 1; It is characterized in that: said drag proofing unit (20) has the mechanical drive (23) that is fixed on middle part, experimental tank chamber (3); This device is made up of direct-current machine (23-1), wheel box (23-2), turn-screw (23-3), driving nut (23-4), portable bar (23-5), axle sleeve (23-6) and overcoat (23-7); On the overcoat (23-7) of its vertical fixing, be fixed with lower fixed frame (20-9) and movable block (20-1) down be housed at the one of which end; On the portable bar (23-5) of mechanical drive (23), be fixed with upper holder (20-8) and movable block (20-2) and micro valve (20-3) be housed at the one of which end; Portable bar (23-5) outside on upper holder (20-8) is with spring (20-7) and on its top gland (20-6) is housed, and between this gland and micro valve (20-3), is provided with fixed briquetting (20-4) and active spill spin block (20-5).
3. the microorganism resistance test apparatus that is used for verification of plasma sterilization according to claim 1 and 2; It is characterized in that: said electrothermic constant temperature equipment (5) is made up of the hot plate and the thermal insulation layer that are located at vacuum tank (1) outside, and is furnished with TP and temperature controller.
4. the microorganism resistance test apparatus that is used for verification of plasma sterilization according to claim 1 and 2; It is characterized in that: said vacuum tank (1) is processed by stainless steel, titanium alloy or duraluminum; Be shaped as drum shape or rectangle, vertical or horizontal placement, volume is 30~150L.
5. the microorganism resistance test apparatus that is used for verification of plasma sterilization according to claim 1 and 2; It is characterized in that: said discharge electrode (4) is processed by stainless steel, titanium alloy or duraluminum porous plate; With the tank skin gap be 5~50mm, and insulate with tank skin.
6. the microorganism resistance test apparatus that is used for verification of plasma sterilization according to claim 1 and 2; It is characterized in that: said neutralizing agent charging point (16) comprises 1~5 syringe and liquid feeding carriage, and each syringe has the substratum that contains neutralizing agent of 5~10ml.
7. the microorganism resistance test apparatus that is used for verification of plasma sterilization according to claim 1 and 2 is characterized in that: said sterile medium charging point (17) comprises syringe and liquid feeding carriage.
8. the microorganism resistance test apparatus that is used for verification of plasma sterilization according to claim 1 and 2 is characterized in that; Near said neutralizing agent charging point (16) and the sterile medium charging point (17) TP I (14-1) is housed.
9. a method of carrying out the microorganism resistance test with the described testing apparatus of claim 2 is characterized in that comprising the steps:
The substratum 10mL that 1) will contain neutralizing agent in 1~5 syringe of suction, is installed in respectively on the liquid feeding carriage one by one, with the Zylox linking conduit I (15-1) of internal diameter 2mm with sealed interface and withstand voltage test tube on syringe, the vacuum tank (1)
(21) rubber seal plug on (22) is connected, and 1~5 test organisms sheet is put into respectively in 1~5 withstand voltage test tube (21), covers rubber seal plug (22);
The withstand voltage test tube (21) that 2) the test organisms sheet will be housed is installed on the lower fixed frame (20-9) of drag proofing unit (20); Clamp with following movable block (20-1); It is fixing with upper holder (20-8) that rubber seal plug (22) is used movable block (20-2), and said Zylox linking conduit I (15-1) is connected with micro valve (20-3);
3) with disinfectant H 2O 2In the syringe of suction sterile medium charging point (17), be installed on the liquid feeding carriage, use internal diameter to be connected on the miniature electromagnetic valve (18) as the tetrafluoroethylene linking conduit II (15-2) of 1.5mm;
4) close the hermatic door (2) of experimental tank chamber (3), on testing apparatus control touch-screen, set 0~999 second grouping time, each bacterial classification is divided into 10 groups according to germicidal action time length and makes an experiment, every group of 1~5 sample;
5) open vacuum pump (7) vacuumizes, and opens the rubber seal plug (22) on the withstand voltage test tube (21);
6) when vacuum tightness reaches 50~150Pa, close the sealing plug of withstand voltage test tube, open plasma exciatiaon power supply (10) and carried out preparatory plasma glow discharge 1~10 minute;
7) open miniature electromagnetic valve (18), add disinfectant H 2O 2And vaporization, keep diffusion in 1~10 minute;
8) sealing plug of opening withstand voltage test tube carries out bactericidal reaction and timing, and the reaction times was controlled at 0~999 second, immediately closed the sealing plug of withstand voltage test tube; Open micro valve (20-3) simultaneously; Make neutralizing agent submergence bacterium sheet, the time control accuracy was 0.1 second, stopped reaction;
9) be evacuated down to 50~150Pa once more, open plasma exciatiaon power supply (10), plasma discharge 1~10 minute;
10) close plasma exciatiaon power supply (10), open inflation interface (24), the inflation concora crush;
11) open hermatic door (2), take out the withstand voltage test tube of band sealing plug;
12) sterilization finishes, and gets 1~3 at random 1~5 bacterium sheet the inside and carries out the enumeration of bacterium sheet.
CN2008101210088A 2008-09-12 2008-09-12 Microorganism resistance test apparatus and method for verification of plasma sterilization Expired - Fee Related CN101363003B (en)

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