One kind resistance bacterium property experimental rig and test methodTechnical 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 andTest 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 hindersThe quality of bacterium performance is particularly important for control trauma surface infestation.However, this is not intended to require that all contact woundsFace 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 performanceContact Wound dressing.In addition, some contact surface of a wound for needing secondary dressing (Secondary Dressing) to use cooperativelyDressing, 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 dressingBreath 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 classesThe 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.OneAs 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 asThe 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 dressingResistance 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 amountGo 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 dressingOuter 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 orVery 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 dressingIt 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 dressingThoroughly evaluating is carried out, at present, the resistance bacterium property also come without suitable experimental rig and test method to Wound dressing carries out completeEvaluate 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 experimentDevice and test method can be to claiming that the resistance bacterium performance of the Wound dressing with resistance bacterium performance is evaluated, and cover dressingDifferent 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 containingThe challenge room of Challenge and method thing and the sampler chamber for cleaning sampling are put, challenge room and sampler chamber combine composition experiment dressPut main body；Test sample is provided between challenge room and sampler chamber, challenge room and sampler chamber clamp test sample；Challenge on roomProvided 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 andSampler 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 roomThe 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 shellBolt, 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 bodyEnd 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 symmetricalArrangement 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 onThe center of sampler chamber sphere.Challenge room and sampler chamber are set to hemispherical, one is that circular section does not have corner, at bothIt 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 toObservation 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 chamberBend 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 rigLoad 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 rigMethod, 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 examinationSample inner surface towards sampler chamber and by test sample vertically be arranged on challenge room and sampler chamber between.To ensure experimental resultAccuracy, generally test intended test 3 samples, test sample takes 3, the result of the test of 3 test samples respectivelyIt 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 examinationThe 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 cutRemainder is wrapped in around experimental rig main body；, can be using the larger sample of area for the less sample of areaRaw 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 propertyDuring 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 equalIt 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 roomIt 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 stateDuring 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 stateDuring 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 externalPollution, 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 propertyDuring experiment, after the completion of culture, moisten sterile swab in advance in sterile saline and squeeze out unnecessary moisture, pass through sampler chamberMouth wipes all inner surfaces of test sample, is then inoculated with the way of wiping TSA culture medium flat plates surface, after inoculationTSA 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 swabThe 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 phaseBetween 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, cultivateMicroorganism 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 resistanceWhen bacterium property is tested, moisten sterile swab in advance in sterile saline and squeeze out unnecessary moisture, by challenging room mouthful wipingTest sample all outer surfaces, are then inoculated with the way of wiping TSA culture medium flat plates surface, and the TSA after inoculation is trainedBase 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 threeIndividual 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 swabWar 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 intoIncubator 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 sampleGrown on the culture medium of product inner surface sampling analysis without microorganism, be judged to meet resistance bacterium property requirement；If for table in sampleThere 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 itBacterium 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 stepSuddenly 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, examinationTest effectively, it is invalid otherwise to test；If given birth on the TSA culture medium flat plates of sample inner surface sampling analysis without microorganismIt 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 analysisWhether 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 haveExtraneous 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 inspectionDuring 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 asThe 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 bacteriumProperty 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 inspectionWhen 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 downGrown 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 downThere 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 notBacterium 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 inspectionWhen, 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 cultivatedBase 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, ifIt 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 bacteriumStrain；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 asUnder (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 boardsUpper 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 stateIt 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 thingSerratia 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 concentrationBacterium 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 dayUse.
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 claimingExperiment, 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 completeThe 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 claimingValency, 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 connectPlant 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 otherEquivalent 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 microorganismMethod 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 boardsUpper 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 stateIt 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 thingSerratia 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 concentrationBacterium 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 dayUse.
Before experiment, sterilizing is carried out to experimental rig using the sterilizing program of confirmed mistake standby.The experiment of following examplesMethod 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 wipingThe 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 samplingTest 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 chosenThe 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 openingIt 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 adoptedIt 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 andBottom 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 tryingExperiment 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 22The 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 sideAngle, is easy to alignment when both combine, while also allowing for batch production；Two be the spherical smooth nothing of experimental rig body inner surfaceTurning, 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 glassMake, 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, willTest 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 occurLeakage, 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 thatTest 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 cutOr 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 areaRow 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 inoculationOnto 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 noExtraneous 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 salineAnd 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 tablesThe 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, oftenIndividual 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 downPlate, 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 manyRemaining 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 surfaceFormula 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 sterileSwab 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 nothingEffect.If grown on the TSA flat boards of sample inner surface sampling analysis without microorganism, being judged to meet dry state resistance bacterium property willAsk.If having microorganism growth on the TSA flat boards for sample inner surface sampling analysis, suitable micro-biological process should be usedWhether 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 huskyThunder 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 huskyThunder 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 chamberA 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 systemInto.
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, willTest 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 occurLeakage, 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 thatTest 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 cutOr 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 areaRow 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 roomMouth 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 mouthBase 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 noExtraneous contamination can be produced, experimental rig is put into 25 DEG C of incubators to 24h, using shaken cultivation case and suitable vibration is setFrequency 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 feelingsCondition.
Step (6)：Challenge microbial activity inspection：Moisten sterile swab 6 in advance in sterile saline and squeeze out manyRemaining 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 surfaceFormula 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 sterileSwab 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 nothingEffect.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 shownWhether muddiness, should use suitable micro-biological process to check it for serratia marcesens.If serratia marcesens, then it is judged to not be inconsistentClose 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 marcesensTSB 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 roomA 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 systemInto.
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, willTest 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 occurLeakage, 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 thatTest 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 cutOr 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 areaRow 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 8Liquid 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 noExtraneous contamination can be produced, experimental rig is put into 25 DEG C of incubators to 24h, using shaken cultivation case and suitable vibration is setFrequency 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 salineAnd 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 tablesThe 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 turnDynamic 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 tryTest invalid.If grown on the TSA flat boards of sample inner surface sampling analysis without microorganism, it is judged to meet half hygrometric state (outsideIt 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 analysisSuitable micro-biological process checks whether it is serratia marcesens.If serratia marcesens, then it is judged to not meet half hygrometric state (exogenous dampIt 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 roomA 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 adoptedIt 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, willTest 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 occurLeakage, 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 thatTest 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 cutOr 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 areaRow 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 8Liquid 32.
Step (3)：Add TSB culture mediums：Appropriate TSB cultures are added into sampler chamber 22 by the sampled room of bend pipe 8 mouthBase 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 noExtraneous contamination can be produced, experimental rig is put into 25 DEG C of incubators to 24h, using shaken cultivation case and suitable vibration is setFrequency 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 confessionThe 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 feelingsCondition.
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 tryTest 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, shouldSuitable micro-biological process is used to check it whether for serratia marcesens.If serratia marcesens, then it is judged to not meet hygrometric stateHinder 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 laboratoryRow 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 limitThe system present invention, within the spirit and principles of the invention, any modification, equivalent substitution and improvements made etc. should be included inWithin 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 protrusionThe innovative characteristicses of the present invention, above-mentioned technical characteristic will not be repeated here.