CN105132272B - One kind resistance bacterium property experimental rig and test method - Google Patents
One kind resistance bacterium property experimental rig and test method Download PDFInfo
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- CN105132272B CN105132272B CN201510560185.6A CN201510560185A CN105132272B CN 105132272 B CN105132272 B CN 105132272B CN 201510560185 A CN201510560185 A CN 201510560185A CN 105132272 B CN105132272 B CN 105132272B
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Abstract
One kind resistance bacterium property experimental rig and test method, described experimental rig includes being used for the challenge room for holding Challenge and method thing and the sampler chamber for cleaning sampling, challenge room and sampler chamber combine composition experimental rig main body, and experimental rig main body is arranged on a support.Described test method includes sample clamping, adds challenge microorganism, sample challenge, penetrates the steps such as microorganism checking, challenge microbial activity inspection and result judgement.The experimental rig and test method of the present invention can be to claiming that the resistance bacterium performance of the Wound dressing with resistance bacterium performance is evaluated, and covers the different conditions of dressing, and evaluation result science is comprehensive.
Description
Technical field
The present invention relates to the resistance bacterium Journal of Sex Research technical field of Wound dressing, specifically a kind of resistance bacterium property experimental rig and
Test method, it is adaptable to claiming that the resistance bacterium performance of the Wound dressing with resistance bacterium performance is evaluated.
Background technology
Clinically, infection is the important complication of wound, and contact Wound dressing is as the mechanical barrier of the surface of a wound, and it hinders
The quality of bacterium performance is particularly important for control trauma surface infestation.However, this is not intended to require that all contact wounds
Face dressing all has resistance bacterium property, and some low-down surface of a wound types of infection risk must not necessarily may be used with resistance bacterium performance
Contact Wound dressing.In addition, some contact surface of a wound for needing secondary dressing (Secondary Dressing) to use cooperatively
Dressing, it hinders bacterium property and provided jointly by both party, only carries out the letter obtained by resistance bacterium property is evaluated to contact Wound dressing
Breath is also likely to be unilateral.Evaluate whether specific contact Wound dressing there should be resistance bacterium performance, it is necessary to different surface of a wound classes
The infection risk of type carries out further investigation.
The inner surface state in which of dressing depend on wound fluid number, correspondingly in hygrometric state or dry state.One
As in home, the outer surface of dressing can be in dry state, and outer surface hygrometric state is not the normality of dressing.But some dressing, such as
The dressing with water preventing ability (such as can be had a bath or be taken a shower) is claimed, its outer surface is also possible to that hygrometric state can be in.To dressing
Resistance bacterium property when being evaluated, residing different conditions should be expected according to dressing and select corresponding test method.
Generally, the state of dressing has following three kinds of situations：
(1) dry state：The two sides of dressing not wet states.Dressing under the state, the surface of a wound oozes without diffusate or very small amount
Go out liquid, and dressing outer surface is in home, it is contemplated that it is not subjected to the processes such as shower；
(2) half hygrometric states：The one side of dressing is wet state.Dressing under the state, the surface of a wound has diffusate to ooze out, and dressing
Outer surface is in home, it is contemplated that be not subjected to the processes such as shower, i.e. " outer dry interior wet "；Or, the surface of a wound without diffusate or
Very small amount diffusate, and dressing outer surface is expected that the processes such as shower can be subjected to, i.e., " done in exogenous damp "；
(3) hygrometric state：The two sides of dressing is all wet state.Dressing under the state, the surface of a wound has diffusate to ooze out, and dressing
It is expected that the processes such as shower can be subjected in outer surface.
The resistance bacterium performance of contact Wound dressing is evaluated, above-mentioned three kinds of states should be taken into full account, to the resistance bacterium performance of dressing
Thoroughly evaluating is carried out, at present, the resistance bacterium property also come without suitable experimental rig and test method to Wound dressing carries out complete
Evaluate in face.
The content of the invention
In view of the above-mentioned problems, it is an object of the invention to provide one kind resistance bacterium property experimental rig and test method, the experiment
Device and test method can be to claiming that the resistance bacterium performance of the Wound dressing with resistance bacterium performance is evaluated, and cover dressing
Different conditions, evaluation result science is comprehensive.
The technical scheme adopted by the invention to solve the technical problem is that：One kind resistance bacterium property experimental rig, including for containing
The challenge room of Challenge and method thing and the sampler chamber for cleaning sampling are put, challenge room and sampler chamber combine composition experiment dress
Put main body；Test sample is provided between challenge room and sampler chamber, challenge room and sampler chamber clamp test sample；Challenge on room
Provided with challenge room mouthful, sampler chamber is challenged provided with sampler chamber mouthful and is equipped with blocking at room mouthful and sampler chamber mouthful.
Further technical scheme is：Described experimental rig main body is arranged on support, described challenge room and
Sampler chamber is in groove-like, and the open side for challenging room and sampler chamber encloses raised edge provided with one along its edge of opening, challenges the convex of room
The raised edge for playing edge and sampler chamber is mutually butted and clamped by the fixing bolt on the support.Pass through raised edge and fixed spiral shell
Bolt, can reliably clamp test sample, so that resistance bacterium property experiment is smoothed out.
Described support includes support frame vertical above horizontal base and base, the top and bottom of experimental rig main body
End is connected by fixing bolt with support frame.
Further technical scheme is：Described challenge room and sampler chamber in hemispherical, challenge room and sampler chamber are symmetrical
Arrangement is so that experimental rig main body challenges the center that room mouth is arranged on challenge room sphere in spherical, and sampler chamber mouthful is arranged on
The center of sampler chamber sphere.Challenge room and sampler chamber are set to hemispherical, one is that circular section does not have corner, at both
It is easy to alignment during combination, while also allowing for batch production；Two be that spherical experimental rig body inner surface is smooth without turning, is easy to
Observation also allows for cleaning.
Further technical scheme is：When carrying out the resistance bacterium property experiment of half hygrometric state, described challenge room mouthful or sampler chamber
Bend pipe is plugged with mouthful；When carrying out hygrometric state resistance bacterium property experiment, bend pipe is plugged with described challenge room mouthful and sampler chamber mouthful；
Described blocking is engaged with the end of bend pipe.When carrying out the experiment of half hygrometric state or hygrometric state, for the ease of in experimental rig
Load challenge bacterium solution or TSB culture mediums in main body, it is necessary on challenge room mouthful or sampler chamber mouthful bridge piece.
Further technical scheme is：Described challenge room, sampler chamber, blocking and bend pipe is made of clear glass.
Above-mentioned part is made using clear glass, is easy to operation and the viewing test result of experiment.
The technical scheme that its technical problem of present invention solution is taken also includes：A kind of experiment side for hindering bacterium property experimental rig
Method, comprises the following steps：
Step (1)：Sample clamping：Test sample is taken, test sample is clamped between challenge room and sampler chamber, is made for examination
Sample inner surface towards sampler chamber and by test sample vertically be arranged on challenge room and sampler chamber between.To ensure experimental result
Accuracy, generally test intended test 3 samples, test sample takes 3, the result of the test of 3 test samples respectively
It is all qualified just qualified.The method of clamping of test sample preferably passes through confirmation, it is ensured that the leakage for challenging microorganism does not occur, will be for examination
The edge of sample is clipped between challenge room and sampler chamber and clamped by fixing bolt.For the sample that area is larger, it can cut
Remainder is wrapped in around experimental rig main body；, can be using the larger sample of area for the less sample of area
Raw material are tested, or lack part can be filled by the way of suitable for testing.
Step (2)：Add challenge microorganism：When the resistance bacterium property experiment of progress dry state or outer dry interior half wet hygrometric state hinder bacterium property
During experiment, smeared with sterile swab and take Challenge and method thing, be inoculated into by challenging room mouthful on the outer surface of test sample, smear equal
It is even；When the resistance bacterium property experiment of half hygrometric state or hygrometric state resistance bacterium property experiment done in exogenous damp, by challenging room mouthful to challenge room
It is middle to add challenge bacterium solution.
Step (3)：Add TSB culture mediums：The experiment of bacterium property or hygrometric state resistance bacterium property are hindered when carrying out outer dry interior half wet hygrometric state
During experiment, TSB culture mediums are added into sampler chamber by sampler chamber mouthful；Hinder what is done in the experiment of bacterium property or exogenous damp when carrying out dry state
During the resistance bacterium property experiment of half hygrometric state, this step is omitted.
Step (4)：Sample is challenged：Room mouthful and sampler chamber mouthful are challenged in closing in an adequate manner, it is ensured that will not be produced external
Pollution, incubator culture is put into by experimental rig.
Step (5)：Penetrate microorganism checking：When half hygrometric state done in dry state resistance bacterium property experiment or exogenous damp hinders bacterium property
During experiment, after the completion of culture, moisten sterile swab in advance in sterile saline and squeeze out unnecessary moisture, pass through sampler chamber
Mouth wipes all inner surfaces of test sample, is then inoculated with the way of wiping TSA culture medium flat plates surface, after inoculation
TSA culture medium flat plates, which are put into incubator culture, is used for sample inner surface sampling analysis.In order to sample fully, also for by swab
The microorganism sampled is fully inoculated on flat board, and each sample is using three sterile swab wipe samples and is inoculated with three times, the phase
Between should repeatedly rotate swab.When carrying out outer dry interior wet half hygrometric state resistance bacterium property experiment or hygrometric state resistance bacterium property experiment, cultivate
Microorganism growing state in Cheng Hou, observation TSB culture mediums.
Step (6)：Challenge microbial activity inspection：When the resistance bacterium property experiment of progress dry state or outer dry interior wet half hygrometric state resistance
When bacterium property is tested, moisten sterile swab in advance in sterile saline and squeeze out unnecessary moisture, by challenging room mouthful wiping
Test sample all outer surfaces, are then inoculated with the way of wiping TSA culture medium flat plates surface, and the TSA after inoculation is trained
Base flat board is supported to be put into incubator culture to challenge the vigor inspection of microorganism.When wiping and being inoculated with, each sample uses three
Individual sterile swab wipe samples and inoculation three times, during which should repeatedly rotate sterile swab.
When the resistance bacterium property experiment of half hygrometric state or hygrometric state resistance bacterium property experiment done in exogenous damp, dipped and chosen with sterile swab
War bacterium solution, is then inoculated with the way of wiping TSA culture medium flat plates surface, the TSA culture medium flat plates after inoculation is put into
Incubator culture for challenge microorganism vigor inspection.
Step (7)：As a result judge：When challenge microorganism is vibrant, experiment is effective, and it is invalid otherwise to test；If for sample
Grown on the culture medium of product inner surface sampling analysis without microorganism, be judged to meet resistance bacterium property requirement；If for table in sample
There is microorganism growth on the culture medium of surface sample analysis, whether be challenge bacterium, if challenge bacterium, then be judged to not meet resistance if checking it
Bacterium property requirement, if not challenge bacterium, then illustrate there is extraneous contamination, it is invalid to test.
Further technical scheme is：When carrying out the resistance bacterium property experiment of half hygrometric state or hygrometric state resistance bacterium property experiment, the step
Suddenly the incubator of (4) is using shaken cultivation case and sets frequency of oscillation, so that challenge bacterium solution fully contacts test sample outer surface,
Or TSB culture mediums is fully contacted test sample inner surface.
Further technical scheme is：The determination methods of the step (7) are：
When carrying out dry state resistance bacterium property experiment, when having challenge bacteria growing on the TSA culture medium flat plates of vigor inspection, examination
Test effectively, it is invalid otherwise to test；If given birth on the TSA culture medium flat plates of sample inner surface sampling analysis without microorganism
It is long, it is judged to meet resistance bacterium property requirement；If having microorganism life on the TSA culture medium flat plates for sample inner surface sampling analysis
Whether long, it is to challenge bacterium to check it, if challenge bacterium, then is judged to not meet resistance bacterium property requirement, if not challenge bacterium, then illustrate have
Extraneous contamination, it is invalid to test.
When carrying out half hygrometric state resistance bacterium property experiment wet in outer do, chosen on the TSA culture medium flat plates of vigor inspection
During war bacteria growing, experiment is effective, and it is invalid otherwise to test；If the display clarification of TSB culture mediums, it is judged to meet resistance bacterium property requirement；Such as
The display of fruit TSB culture mediums is muddy, and whether be challenge bacterium, if challenge bacterium if checking the microorganism in it, then be judged to not meet resistance bacterium
Property require, if not challenge bacterium, then explanation has an extraneous contamination, and it is invalid to test.
When the half hygrometric state resistance bacterium property experiment done in exogenous damp, chosen on the TSA culture medium flat plates of vigor inspection
When bacteria suspension of fighting grows, experiment is effective, and it is invalid otherwise to test；If the TSA culture mediums for sample inner surface sampling analysis are put down
Grown on plate without microorganism, be judged to meet resistance bacterium property requirement；If the TSA culture mediums for sample inner surface sampling analysis are put down
There is microorganism growth on plate, whether be challenge bacterium, if challenge bacterium if checking it, be then judged to not meet resistance bacterium property requirement, if not
Bacterium is challenged, then explanation has extraneous contamination, it is invalid to test.
When carrying out hygrometric state resistance bacterium property experiment, there is challenge bacteria suspension growth on the TSA culture medium flat plates of vigor inspection
When, experiment is effective, and it is invalid otherwise to test；If the display clarification of TSB culture mediums, it is judged to meet resistance bacterium property requirement；If TSB is cultivated
Base display is muddy, and whether be challenge bacterium, if challenge bacterium if checking the microorganism in it, then is judged to not meet resistance bacterium property requirement, if
It is not challenge bacterium, then explanation has extraneous contamination, and it is invalid to test.
The reagent and material that the test method of the present invention is used have：Serratia marcesens, ATCC 8100, or other equivalent bacterium
Strain；Trypticase soya broth (TSB)；Tryptic Soy Agar (TSA)；Sterile saline；Sterile swab.
Before being tested, challenge microorganism should be prepared, test method challenge microorganism preparation method of the invention is such as
Under (by taking serratia marcesens as an example)：
It is prepared by Challenge and method thing：From picking serratia marcesens bacterium colony on serratia marcesens work bacterial strain inclined-plane, in TSA flat boards
Upper streak inoculation, Challenge and method thing is made in incubated overnight under the conditions of 25 DEG C, (outer dry for dry state resistance bacterium property experiment and half hygrometric state
It is interior wet) resistance bacterium property experiment.The flat board prepared is before use in 4 DEG C of preservations, and the same day uses.
Bacterium solution is challenged to prepare：Picking on the obtained TSA flat boards with Challenge and method thing is prepared from above-mentioned Challenge and method thing
Serratia marcesens single bacterium colony is inoculated in TSB culture mediums, incubated overnight under the conditions of 25 DEG C, and it is about 109cfu/mL's to obtain concentration
Bacterium solution is challenged, for half hygrometric state (exogenous damp is dry interior) resistance bacterium property experiment.The challenge bacterium solution prepared is before use in 4 DEG C of preservations, the same day
Use.
The beneficial effects of the invention are as follows：
1st, there is provided a kind of experimental rig, for having the Wound dressing of resistance bacterium performance to carry out resistance bacterium performance evaluation to claiming
Experiment, can carry out the experiment of the different conditions such as dry state, half hygrometric state and hygrometric state, and the resistance bacterium performance progress science to Wound dressing is complete
The evaluation in face；
2nd, there is provided a kind of test method, commented for having the Wound dressing of resistance bacterium performance to carry out resistance bacterium performance to claiming
Valency, and resistance bacterium performance evaluation of the dressing under the different conditions such as dry state, half hygrometric state, hygrometric state is covered, evaluation result science is comprehensive.
Brief description of the drawings
Fig. 1 is carrying out structural representation when dry state resistance bacterium property is tested for the experimental rig of the present invention；
Fig. 2 is structural representation of the experimental rig of the present invention when carrying out half hygrometric state (outer dry interior wet) resistance bacterium property experiment；
Fig. 3 is carrying out the structural representation half hygrometric state (does) resistance bacterium property experiment in exogenous damp when for the experimental rig of the present invention；
Fig. 4 is carrying out structural representation when hygrometric state resistance bacterium property is tested for the experimental rig of the present invention.
In figure：1 blocks, 2 experimental rig main bodys, and 21 challenge rooms, 22 sampler chambers, 3 Challenge and method things, 31 serratia marcesens connect
Plant thing, 32 challenge bacterium solutions, 4 fixing bolts, 5 test samples, 6 sterile swabs, 7 supports, 8 bend pipes, 9TSB culture mediums.
Embodiment
With reference to Figure of description and specific embodiment, the invention will be further described：
The reagent and material that the test method of the embodiment of the present invention is used have：Serratia marcesens, ATCC 8100, or it is other
Equivalent bacterial strain；Trypticase soya broth (TSB)；Tryptic Soy Agar (TSA)；Sterile saline；Sterile swab.
Before being tested, challenge microorganism should be prepared, the test method of the embodiment of the present invention challenges the preparation of microorganism
Method is following (by taking serratia marcesens as an example)：
It is prepared by Challenge and method thing：From picking serratia marcesens bacterium colony on serratia marcesens work bacterial strain inclined-plane, in TSA flat boards
Upper streak inoculation, Challenge and method thing is made in incubated overnight under the conditions of 25 DEG C, (outer dry for dry state resistance bacterium property experiment and half hygrometric state
It is interior wet) resistance bacterium property experiment.The flat board prepared is before use in 4 DEG C of preservations, and the same day uses.
Bacterium solution is challenged to prepare：Picking on the obtained TSA flat boards with Challenge and method thing is prepared from above-mentioned Challenge and method thing
Serratia marcesens single bacterium colony is inoculated in TSB culture mediums, incubated overnight under the conditions of 25 DEG C, and it is about 109cfu/mL's to obtain concentration
Bacterium solution is challenged, for half hygrometric state (exogenous damp is dry interior) resistance bacterium property experiment.The challenge bacterium solution prepared is before use in 4 DEG C of preservations, the same day
Use.
Before experiment, sterilizing is carried out to experimental rig using the sterilizing program of confirmed mistake standby.The experiment of following examples
Method is used as challenge bacterium using serratia marcesens.
Embodiment one：
As shown in Figure 1.One kind resistance bacterium property experimental rig, including for holding the challenge room 21 of Challenge and method thing and for wiping
The sampler chamber 22 of sampling is wiped, challenge room 21 and sampler chamber 22 combine composition experimental rig main body 2, challenge room 21 and sampling
Test sample 5 is provided between room 22, challenge room 21 and sampler chamber 22 clamp test sample 5.
Experimental rig main body 2 is arranged on a support 7；Described challenge room 21 and sampler chamber 22 are in groove-like, are chosen
The open side of war room 21 and sampler chamber 22, provided with the raised edge of a circle, challenges raised edge and the sampler chamber 22 of room 21 along its edge of opening
It is raised along being mutually butted and clamped by the fixing bolt 4 on the support 7；Challenge room mouthful is additionally provided with challenge room 21, is adopted
It is additionally provided with specimen chamber 22 at sampler chamber mouthful, challenge room mouthful and sampler chamber mouthful and is equipped with blocking 1.
Described support 7 includes support frame vertical above horizontal base and base, the top of experimental rig main body 2 and
Bottom is connected by fixing bolt 4 with support frame.
Described challenge room 21 and sampler chamber 22 in hemispherical, challenge room 21 and sampler chamber 22 are arranged symmetrically so that trying
Experiment device main body 2 challenges the center that room mouth is arranged on the challenge sphere of room 21 in spherical, and sampler chamber mouthful is arranged on the ball of sampler chamber 22
The center in face.Challenge room 21 and sampler chamber 22 are set to hemispherical, had the following advantages：One is that circular section does not have side
Angle, is easy to alignment when both combine, while also allowing for batch production；Two be the spherical smooth nothing of experimental rig body inner surface
Turning, is easy to observation to also allow for cleaning.
Described challenge room 21, sampler chamber 22, block 1 and be made of clear glass.Above-mentioned part uses clear glass
Make, be easy to operation and the viewing test result of experiment.
The experimental rig of the present embodiment is used to carry out dry state resistance bacterium property experiment.
The test method (experiment of dry state resistance bacterium property) of the experimental rig of the present embodiment comprises the following steps：
Step (1)：Sample clamping：3 test samples 5 are taken respectively, make the inner surface of test sample 5 towards sampler chamber 22, will
Test sample 5 is clamped between challenge room 21 and sampler chamber 22.Method of clamping preferably passes through confirmation, it is ensured that challenge microorganism does not occur
Leakage, by the edge of test sample 5 be clipped in challenge room 21 and sampler chamber 22 between and clamped by fixing bolt 4 so that
Test sample 5 is vertical to be arranged between challenge room 21 and sampler chamber 22.For the sample that area is larger, remainder can be cut
Or be wrapped in around experimental rig main body 2；For the less sample of area, it can be entered using the larger sample raw material of area
Row test, or lack part can be filled by the way of suitable for testing.
Step (2)：Add challenge microorganism：Smeared with sterile swab 6 and take Challenge and method thing 3, and by challenging room mouthful inoculation
Onto the outer surface of test sample 5, smear as far as possible uniform.
Step (3)：Sample is challenged：The challenge room mouthful of blocking test device and sampler chamber mouthful in an adequate manner, it is ensured that no
Extraneous contamination can be produced, experimental rig is put into 25 DEG C of incubators to 24h.
Step (4)：Penetrate microorganism checking：To after the stipulated time, sterile swab 6 is moistened in advance in sterile saline
And squeeze out unnecessary moisture, all inner surfaces of test sample 5 are wiped by sampler chamber mouthful as far as possible, then to wipe TSA flat board tables
The mode in face is inoculated with.In order to sample fully, fully it is inoculated into also for the microorganism for being upsampled to swab on flat board, often
Individual test sample 5 is using three sterile swab wipe samples and is inoculated with three times, during which should repeatedly rotate sterile swab 6.TSA is put down
Plate, which is placed in 35 DEG C of culture 24h, is used for sample inner surface sampling analysis.
Step (5)：Challenge microbial activity inspection：Moisten sterile swab 6 in advance in sterile saline and squeeze out many
Remaining moisture, all outer surfaces of test sample 5 are wiped by challenging room mouthful, then to wipe the side on TSA culture medium flat plates surface
Formula is inoculated with, and each test sample 5 is using three sterile swab wipe samples and is inoculated with three times, during which should repeatedly rotate sterile
Swab 6.TSA culture medium flat plates after inoculation are placed in the vigor inspection that 35 DEG C of culture 24h are used to challenge microorganism.
Step (6)：As a result judge：There should be serratia marcesens growth on the TSA flat boards of vigor inspection, otherwise test nothing
Effect.If grown on the TSA flat boards of sample inner surface sampling analysis without microorganism, being judged to meet dry state resistance bacterium property will
Ask.If having microorganism growth on the TSA flat boards for sample inner surface sampling analysis, suitable micro-biological process should be used
Whether check it is serratia marcesens.If serratia marcesens, then it is judged to not meet dry state resistance bacterium property requirement；If not cement is husky
Thunder bacterium, then illustrate there is extraneous contamination, it is invalid to test.
Serratia marcesens can show red colonies in agar surface, and this feature can be used for differentiating whether have cement husky
Thunder bacteria growing.
Embodiment two：
As shown in Figure 2.The experimental rig of the present embodiment and the experimental rig difference of embodiment one are：Described sampler chamber
A bend pipe 8 is plugged with mouthful, described blocking 1 is engaged with the end of bend pipe 8, described bend pipe 8 also uses clear glass system
Into.
The experimental rig of the present embodiment is used to carry out half hygrometric state (outer dry interior wet) resistance bacterium property experiment.
The test method [half hygrometric state (outer dry interior wet) resistance bacterium property experiment] of the experimental rig of the present embodiment comprises the following steps：
Step (1)：Sample clamping：3 test samples 5 are taken respectively, make the inner surface of test sample 5 towards sampler chamber 22, will
Test sample 5 is clamped between challenge room 21 and sampler chamber 22.Method of clamping preferably passes through confirmation, it is ensured that challenge microorganism does not occur
Leakage, by the edge of test sample 5 be clipped in challenge room 21 and sampler chamber 22 between and clamped by fixing bolt 4 so that
Test sample 5 is vertical to be arranged between challenge room 21 and sampler chamber 22.For the sample that area is larger, remainder can be cut
Or be wrapped in around experimental rig main body 2；For the less sample of area, it can be entered using the larger sample raw material of area
Row test, or lack part can be filled by the way of suitable for testing.
Step (2)：Add challenge microorganism：Smeared with sterile swab 6 and take serratia marcesens inoculum 31, and by challenging room
Mouth is inoculated on the outer surface of test sample 5, smears uniform as far as possible.
Step (3)：Add TSB culture mediums：Appropriate TSB cultures are added into sampler chamber 22 by the sampled room of bend pipe 8 mouth
Base 9.
Step (4)：Sample is challenged：The challenge room mouthful of blocking test device and sampler chamber mouthful in an adequate manner, it is ensured that no
Extraneous contamination can be produced, experimental rig is put into 25 DEG C of incubators to 24h, using shaken cultivation case and suitable vibration is set
Frequency is so that TSB culture mediums 9 can fully contact the inner surface of test sample 5.
Step (5)：Penetrate microorganism checking：To after the stipulated time, visually observe microorganism in TSB culture mediums 9 and grow feelings
Condition.
Step (6)：Challenge microbial activity inspection：Moisten sterile swab 6 in advance in sterile saline and squeeze out many
Remaining moisture, all outer surfaces of test sample 5 are wiped by challenging room mouthful, then to wipe the side on TSA culture medium flat plates surface
Formula is inoculated with, and each test sample 5 is using three sterile swab wipe samples and is inoculated with three times, during which should repeatedly rotate sterile
Swab 6.TSA culture medium flat plates after inoculation are placed in the vigor inspection that 35 DEG C of culture 24h are used to challenge microorganism.
Step (7)：As a result judge：There should be serratia marcesens growth on the TSA flat boards of vigor inspection, otherwise test nothing
Effect.If the display clarification of TSB culture mediums, it is judged to meet half hygrometric state (outer dry interior wet) resistance bacterium property requirement.If TSB culture mediums are shown
Whether muddiness, should use suitable micro-biological process to check it for serratia marcesens.If serratia marcesens, then it is judged to not be inconsistent
Close half hygrometric state (outer dry interior wet) resistance bacterium property requirement；If not serratia marcesens, then illustrate there is extraneous contamination, it is invalid to test.
If it is necessary, sterile TSB culture mediums can be set as negative control (display is clarified) and inoculation serratia marcesens
TSB culture mediums as positive control (display muddy), to avoid visually observing clarification or muddy possible operative error.
Embodiment three：
As shown in Figure 3.The experimental rig of the present embodiment and the experimental rig difference of embodiment one are：Described challenge room
A bend pipe 8 is plugged with mouthful, described blocking 1 is engaged with the end of bend pipe 8, described bend pipe 8 also uses clear glass system
Into.
The experimental rig of the present embodiment is used to carry out half hygrometric state (dry in exogenous damp) resistance bacterium property experiment.
The test method [half hygrometric state (does) experiment of resistance bacterium property in exogenous damp] of the experimental rig of the present embodiment comprises the following steps：
Step (1)：Sample clamping：3 test samples 5 are taken respectively, make the inner surface of test sample 5 towards sampler chamber 22, will
Test sample 5 is clamped between challenge room 21 and sampler chamber 22.Method of clamping preferably passes through confirmation, it is ensured that challenge microorganism does not occur
Leakage, by the edge of test sample 5 be clipped in challenge room 21 and sampler chamber 22 between and clamped by fixing bolt 4 so that
Test sample 5 is vertical to be arranged between challenge room 21 and sampler chamber 22.For the sample that area is larger, remainder can be cut
Or be wrapped in around experimental rig main body 2；For the less sample of area, it can be entered using the larger sample raw material of area
Row test, or lack part can be filled by the way of suitable for testing.
Step (2)：Add challenge microorganism：Appropriate challenge bacterium is added into challenge room 21 through challenging room mouthful by bend pipe 8
Liquid 32.
Step (3)：Sample is challenged：The challenge room mouthful of blocking test device and sampler chamber mouthful in an adequate manner, it is ensured that no
Extraneous contamination can be produced, experimental rig is put into 25 DEG C of incubators to 24h, using shaken cultivation case and suitable vibration is set
Frequency is so that challenge bacterium solution 32 can fully contact the outer surface of test sample 5.
Step (4)：Penetrate microorganism checking：To after the stipulated time, sterile swab 6 is moistened in advance in sterile saline
And squeeze out unnecessary moisture, all inner surfaces of test sample 5 are wiped by sampler chamber mouthful as far as possible, then to wipe TSA flat board tables
The mode in face is inoculated with.Each test sample 5 is using three sterile swab wipe samples and is inoculated with three times, during which should repeatedly turn
Dynamic sterile swab 6.TSA flat boards are placed in into 35 DEG C of culture 24h is used for sample inner surface sampling analysis.
Step (5)：Challenge microbial activity inspection：Challenge bacterium solution 32, which is dipped, with sterile swab 6 wipes inoculation TSA flat boards,
TSA culture medium flat plates after inoculation are placed in the vigor inspection that 35 DEG C of culture 24h are used to challenge microorganism.
Step (6)：As a result judge：There should be the growth of serratia marcesens suspension on the TSA flat boards of vigor inspection, otherwise try
Test invalid.If grown on the TSA flat boards of sample inner surface sampling analysis without microorganism, it is judged to meet half hygrometric state (outside
It is wet interior dry) resistance bacterium property requirement., should be using suitable if having microorganism growth on the TSA flat boards for sample inner surface sampling analysis
Suitable micro-biological process checks whether it is serratia marcesens.If serratia marcesens, then it is judged to not meet half hygrometric state (exogenous damp
It is interior dry) resistance bacterium property requirement；If not serratia marcesens, then illustrate there is extraneous contamination, it is invalid to test.
Example IV：
As shown in Figure 4.The experimental rig of the present embodiment and the experimental rig difference of embodiment one are：Described challenge room
A bend pipe 8 is plugged with mouth and sampler chamber mouthful, described blocking 1 is engaged with the end of bend pipe 8, and described bend pipe 8 is also adopted
It is made of clear glass.
The experimental rig of the present embodiment is used to carry out hygrometric state resistance bacterium property experiment.
The test method (experiment of hygrometric state resistance bacterium property) of the experimental rig of the present embodiment comprises the following steps：
Step (1)：Sample clamping：3 test samples 5 are taken respectively, make the inner surface of test sample 5 towards sampler chamber 22, will
Test sample 5 is clamped between challenge room 21 and sampler chamber 22.Method of clamping preferably passes through confirmation, it is ensured that challenge microorganism does not occur
Leakage, by the edge of test sample 5 be clipped in challenge room 21 and sampler chamber 22 between and clamped by fixing bolt 4 so that
Test sample 5 is vertical to be arranged between challenge room 21 and sampler chamber 22.For the sample that area is larger, remainder can be cut
Or be wrapped in around experimental rig main body 2；For the less sample of area, it can be entered using the larger sample raw material of area
Row test, or lack part can be filled by the way of suitable for testing.
Step (2)：Add challenge microorganism：Appropriate challenge bacterium is added into challenge room 21 through challenging room mouthful by bend pipe 8
Liquid 32.
Step (3)：Add TSB culture mediums：Appropriate TSB cultures are added into sampler chamber 22 by the sampled room of bend pipe 8 mouth
Base 9.
Step (4)：Sample is challenged：The challenge room mouthful of blocking test device and sampler chamber mouthful in an adequate manner, it is ensured that no
Extraneous contamination can be produced, experimental rig is put into 25 DEG C of incubators to 24h, using shaken cultivation case and suitable vibration is set
Frequency is so that challenge bacterium solution 32 can fully contact the outer surface of test sample 5, while alloing TSB culture mediums 9 fully to contact confession
The inner surface of test agent 5.
Step (5)：Penetrate microorganism checking：To after the stipulated time, visually observe microorganism in TSB culture mediums 9 and grow feelings
Condition.
Step (6)：Challenge microbial activity inspection：Challenge bacterium solution 32, which is dipped, with sterile swab 6 wipes inoculation TSA flat boards,
TSA culture medium flat plates after inoculation are placed in the vigor inspection that 35 DEG C of culture 24h are used to challenge microorganism.
Step (7) result judges：There should be the growth of serratia marcesens suspension on the TSA flat boards of vigor inspection, otherwise try
Test invalid.If the display clarification of TSB culture mediums, it is judged to meet hygrometric state resistance bacterium property requirement.If the display of TSB culture mediums is muddy, should
Suitable micro-biological process is used to check it whether for serratia marcesens.If serratia marcesens, then it is judged to not meet hygrometric state
Hinder the requirement of bacterium property；If not serratia marcesens, then illustrate there is extraneous contamination, it is invalid to test.
The test method of the present invention needs to operate microorganism, is preferably entered by trained personnel in biocontainment laboratory
Row experiment.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not the whole embodiments of the present invention, not to limit
The system present invention, within the spirit and principles of the invention, any modification, equivalent substitution and improvements made etc. should be included in
Within protection scope of the present invention.
In addition to technical characteristic described in specification, remaining technical characteristic is technology known to those skilled in the art, for protrusion
The innovative characteristicses of the present invention, above-mentioned technical characteristic will not be repeated here.
Claims (5)
1. one kind resistance bacterium property experimental rig, it is characterized in that, including for holding the challenge room of Challenge and method thing and being adopted for wiping
The sampler chamber of sample, challenge room and sampler chamber combine composition experimental rig main body；It is provided between challenge room and sampler chamber
Test sample, challenge room and sampler chamber clamp test sample；Room is challenged provided with challenge room mouthful, sampler chamber is provided with sampler chamber
Mouthful, it is equipped with blocking at challenge room mouthful and sampler chamber mouthful；
Described experimental rig main body is arranged on a support, and described challenge room and sampler chamber are in groove-like, challenge room
With the open side of sampler chamber along its edge of opening provided with the raised edge of a circle, challenge room raised edge and sampler chamber it is raised along mutual
Dock and clamped by the fixing bolt on the support；
Described support includes support frame vertical above horizontal base and base, and the top and bottom of experimental rig main body lead to
Fixing bolt is crossed with support frame to be connected；
Described challenge room and sampler chamber in hemispherical, challenge room and sampler chambers is arranged symmetrically so that experimental rig main body is in
Spherical, challenge room mouth is arranged on the center of challenge room sphere, and sampler chamber mouth is arranged on the center of sampler chamber sphere；
When carrying out the resistance bacterium property experiment of half hygrometric state, bend pipe is plugged with described challenge room mouthful or sampler chamber mouthful；It is wet when carrying out
During state resistance bacterium property experiment, bend pipe is plugged with described challenge room mouthful and sampler chamber mouthful；Described blocking and the end of bend pipe
It is engaged.
2. a kind of resistance bacterium property experimental rig according to claim 1, it is characterized in that, described challenge room, sampler chamber, blocking
It is made with bend pipe of clear glass.
3. based on a kind of test method of resistance bacterium property experimental rig described in claim 1, it is characterized in that, comprise the following steps：
Step (1)：Sample clamping：Test sample is taken, test sample is clamped between challenge room and sampler chamber, makes test sample
Inner surface towards sampler chamber and by test sample vertically be arranged on challenge room and sampler chamber between；
Step (2)：Add challenge microorganism：When the resistance bacterium property experiment of progress dry state or outer dry interior wet half hygrometric state resistance bacterium property experiment
When, smeared with sterile swab and take Challenge and method thing, be inoculated into by challenging room mouthful on the outer surface of test sample, smear uniform；When
When carrying out the resistance bacterium property experiment of half hygrometric state or hygrometric state resistance bacterium property experiment done in exogenous damp, added by challenging room mouthful into challenge room
Challenge bacterium solution；
Step (3)：Add TSB culture mediums：The experiment of bacterium property or hygrometric state resistance bacterium property experiment are hindered when carrying out outer dry interior half wet hygrometric state
When, TSB culture mediums are added into sampler chamber by sampler chamber mouthful；It is semi-moist when what is done in the resistance bacterium property experiment of progress dry state or exogenous damp
During state resistance bacterium property experiment, this step is omitted；
Step (4)：Sample is challenged：Closing challenge room mouthful and sampler chamber mouthful, incubator culture is put into by experimental rig；
Step (5)：Penetrate microorganism checking：When the half hygrometric state resistance bacterium property experiment done in dry state resistance bacterium property experiment or exogenous damp
When, after the completion of culture, moisten sterile swab in advance in sterile saline and squeeze out unnecessary moisture, pass through sampler chamber mouthful and wipe
All inner surfaces of test sample are wiped, are then inoculated with the way of wiping TSA culture medium flat plates surface, by the TSA after inoculation
Culture medium flat plate, which is put into incubator culture, is used for sample inner surface sampling analysis；The examination of bacterium property is hindered when carrying out outer dry interior half wet hygrometric state
Test or hygrometric state resistance bacterium property experiment when, after the completion of culture, observation TSB culture mediums in microorganism growing state；
Step (6)：Challenge microbial activity inspection：When the resistance bacterium property experiment of progress dry state or outer dry interior half wet hygrometric state hinder bacterium property
During experiment, moisten sterile swab in advance in sterile saline and squeeze out unnecessary moisture, wiped by challenging room mouthful for examination
Sample all outer surfaces, are then inoculated with the way of wiping TSA culture medium flat plates surface, by the TSA culture mediums after inoculation
Flat board is put into incubator culture to challenge the vigor inspection of microorganism；
When the resistance bacterium property experiment of half hygrometric state or hygrometric state resistance bacterium property experiment done in exogenous damp, challenge bacterium is dipped with sterile swab
Liquid, is then inoculated with the way of wiping TSA culture medium flat plates surface, and the TSA culture medium flat plates after inoculation are put into culture
Case culture for challenge microorganism vigor inspection；
Step (7)：As a result judge：When challenge microorganism is vibrant, experiment is effective, and it is invalid otherwise to test；If in sample
Grown on the culture medium of surface sampling analysis without microorganism, be judged to meet resistance bacterium property requirement；If adopted for sample inner surface
There is microorganism growth on the culture medium of sample analysis, whether be challenge bacterium, if challenge bacterium if checking it, then be judged to not meet resistance bacterium property
It is required that, if not challenge bacterium, then illustrate there is extraneous contamination, it is invalid to test.
4. a kind of test method for hindering bacterium property experimental rig according to claim 3, it is characterized in that, when half hygrometric state resistance of progress
When bacterium property is tested or hygrometric state resistance bacterium property is tested, the incubator of the step (4) uses shaken cultivation case and sets frequency of oscillation,
So that challenge bacterium solution fully contacts test sample outer surface, or TSB culture mediums are made fully to contact test sample inner surface.
5. a kind of test method of resistance bacterium property experimental rig according to claim 3 or 4, it is characterized in that, the step (7)
Determination methods be：
When carrying out dry state resistance bacterium property experiment, when having challenge bacteria growing on the TSA culture medium flat plates of vigor inspection, experiment has
Effect, it is invalid otherwise to test；If grown on the TSA culture medium flat plates of sample inner surface sampling analysis without microorganism, sentence
To meet resistance bacterium property requirement；If there is microorganism growth on the TSA culture medium flat plates for sample inner surface sampling analysis, check
Whether it is challenge bacterium, if challenge bacterium, then is judged to not meet resistance bacterium property requirement, if not challenging bacterium, then explanation has external dirt
Dye, it is invalid to test；When carrying out half hygrometric state resistance bacterium property experiment wet in outer do, the TSA culture medium flat plates checked for vigor
When having challenge bacteria growing, experiment is effective, and it is invalid otherwise to test；If the display clarification of TSB culture mediums, being judged to meet resistance bacterium property will
Ask；If the display of TSB culture mediums is muddy, whether be challenge bacterium, if challenge bacterium, then be judged to not meet if checking the microorganism in it
The requirement of bacterium property is hindered, if not challenge bacterium, then illustrate there is extraneous contamination, it is invalid to test；
When the half hygrometric state resistance bacterium property experiment done in exogenous damp, there is challenge bacterium on the TSA culture medium flat plates of vigor inspection
When suspension grows, experiment is effective, and it is invalid otherwise to test；If the TSA culture medium flat plates for sample inner surface sampling analysis
Grown without microorganism, be judged to meet resistance bacterium property requirement；If the TSA culture medium flat plates for sample inner surface sampling analysis
There is microorganism growth, whether be challenge bacterium, if challenge bacterium if checking it, be then judged to not meet resistance bacterium property requirement, if not challenge
Bacterium, then illustrate there is extraneous contamination, it is invalid to test；
When carrying out hygrometric state resistance bacterium property experiment, when having challenge bacteria suspension growth on the TSA culture medium flat plates of vigor inspection, examination
Test effectively, it is invalid otherwise to test；If the display clarification of TSB culture mediums, it is judged to meet resistance bacterium property requirement；If TSB culture mediums show
Show muddiness, whether be challenge bacterium, if challenge bacterium if checking the microorganism in it, be then judged to not meet resistance bacterium property requirement, if not
Bacterium is challenged, then explanation has extraneous contamination, it is invalid to test.
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| CN101504354A (en) * | 2009-03-18 | 2009-08-12 | 中国科学院武汉岩土力学研究所 | Impervious material permeation deformation test apparatus for refuse landfill pad |
| WO2009149203A1 (en) * | 2008-06-04 | 2009-12-10 | Kci Licensing, Inc. | Detecting infection in reduced pressure wound treatment |
| CN202011880U (en) * | 2011-01-10 | 2011-10-19 | 山东新华医疗器械股份有限公司 | Tester for bacteria blocking property of gas permeable packing material in aseptic apparatus |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2009149203A1 (en) * | 2008-06-04 | 2009-12-10 | Kci Licensing, Inc. | Detecting infection in reduced pressure wound treatment |
| CN101504354A (en) * | 2009-03-18 | 2009-08-12 | 中国科学院武汉岩土力学研究所 | Impervious material permeation deformation test apparatus for refuse landfill pad |
| CN202011880U (en) * | 2011-01-10 | 2011-10-19 | 山东新华医疗器械股份有限公司 | Tester for bacteria blocking property of gas permeable packing material in aseptic apparatus |
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