A kind of resistance bacterium property testing apparatus 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 testing apparatus and test method, being applicable 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 the quality of its resistance bacterium performance seems particularly important for control traumatic infection.But this does not also mean that requiring that all contact Wound dressings all have hinders bacterium property, and the low-down surface of a wound type of some infection risk may not use the contact Wound dressing with resistance bacterium performance.In addition, some need secondary dressing (SecondaryDressing) with the use of contact Wound dressing, its resistance bacterium property provides by both party jointly, and it also may be unilateral for only carrying out to contact Wound dressing the information that resistance bacterium property evaluation obtains.Evaluate specific contact Wound dressing and whether should have resistance bacterium performance, need to carry out further investigation to the infection risk of different surface of a wound type.
State residing for the internal surface of dressing depends on the number of wound fluid, is correspondingly in hygrometric state or dry state.Normally in environment, the outside surface of dressing can be in dry state, and outside surface hygrometric state is not the normality of dressing.But some dressing, have the dressing of water preventing ability (as carried out having a bath or shower) as claimed, its outside surface also may be in hygrometric state.When evaluating the resistance bacterium property of dressing, corresponding test method should be selected by the different states residing for dressing expection.
Generally, the state of dressing has following three kinds of situations:
(1) dry state: the state that the two sides of dressing is not wet.Dressing under this state, the surface of a wound is without transudate or minute quantity transudate, and dressing outer surface is in home, expects without undergoing processes such as such as shower;
(2) half hygrometric states: the one side of dressing is wet state.Dressing under this state, the surface of a wound has transudate to ooze out, and dressing outer surface is in home, expects without undergoing processes such as such as shower, i.e. " outer dry interior wet "; Or the surface of a wound is without transudate or minute quantity transudate, and the expection of dressing outside surface can stand the processes such as such as shower, i.e. " dry in exogenous damp ";
(3) hygrometric state: the two sides of dressing is all wet state.Dressing under this state, the surface of a wound has transudate to ooze out, and the expection of dressing outside surface can stand the processes such as such as shower.
Evaluate the resistance bacterium performance of contact Wound dressing, above-mentioned three kinds of states should be taken into full account, thoroughly evaluating is carried out to the resistance bacterium performance of dressing at present, also do not have suitable testing apparatus and the resistance bacterium property of test method to Wound dressing to carry out thoroughly evaluating.
Summary of the invention
For the problems referred to above, the object of the present invention is to provide a kind of resistance bacterium property testing apparatus and test method, this testing apparatus and test method to claiming that the resistance bacterium performance of the Wound dressing with resistance bacterium performance is evaluated, and can cover the different states of dressing, and evaluation result science is comprehensive.
The present invention solves the technical scheme that its technical problem takes: a kind of resistance bacterium property testing apparatus, and comprise for holding the challenge room of Challenge and method thing and the sampler chamber for cleaning sampling, challenge room and sampler chamber are combined and formed testing apparatus main body; Be provided with test sample between challenge room and sampler chamber, test sample clamps by challenge room and sampler chamber; Challenge room is provided with challenge room mouth, and sampler chamber is provided with sampler chamber mouth, and challenge room mouth and sampler chamber mouth place are equipped with blocking.
Further technical scheme is: described testing apparatus main body is arranged on a support, described challenge room and sampler chamber are all in groove-like, the open side of challenge room and sampler chamber is provided with the protruding edge of a circle along its edge of opening, and the projection edge of challenge room and the projection of sampler chamber are along mutually docking and passing through the standing bolt clamping on described support.By protruding edge and standing bolt, test sample can be clamped reliably, test to make resistance bacterium property and carry out smoothly.
Described support comprises bracing frame vertical above the base of level and base, and the top of testing apparatus main body is connected with bracing frame by standing bolt with bottom.
Further technical scheme is: described challenge room and sampler chamber are all in hemispherical, challenge room and sampler chamber are arranged symmetrically with thus make testing apparatus main body be 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.Be set to hemispherical by challenge room and sampler chamber, one is that circular cross section does not have corner, is convenient to alignment, is also convenient to batch production simultaneously when both combine; Two is that spherical testing apparatus body inner surface is smooth without turning, is convenient to observe and is also convenient to cleaning.
Further technical scheme is: when carrying out half hygrometric state resistance bacterium property test, described challenge room mouth or sampler chamber mouth are plugged with bend pipe; When carrying out hygrometric state resistance bacterium property test, described challenge room mouth and sampler chamber mouth are all plugged with bend pipe; Described blocking matches with the end of bend pipe.When carrying out the test of half hygrometric state or hygrometric state, for the ease of loading challenge bacterium liquid or TSB substratum in testing apparatus main body, need bridge piece on challenge room mouth or sampler chamber mouth.
Further technical scheme is: described challenge room, sampler chamber, blocking and bend pipe all adopt transparent glass to make.Above-mentioned parts adopt transparent glass to make, and are convenient to operation and the viewing test result of test.
The present invention solves the technical scheme that its technical problem takes and also comprises: a kind of test method hindering bacterium property testing apparatus, comprises the following steps:
Step (1): sample clamping: get test sample, is clamped to test sample between challenge room and sampler chamber, makes test sample internal surface towards sampler chamber and is arranged between challenge room and sampler chamber by vertical for test sample.For ensureing the accuracy of experimental result, generally test intended tests 3 samples, and test sample gets 3 respectively, and the test-results of 3 test samples is all qualified just qualified.The method of clamping of test sample should pass through confirmation, guarantee not occur the leakage of challenging microorganism, the edge clamping of test sample is clamped by standing bolt between challenge room and sampler chamber.For the sample that area is larger, can remainder be cut or be wrapped in around testing apparatus main body; For the sample that area is less, the sample starting material that can area be adopted larger are tested, or suitable mode can be adopted to fill lack part for test.
Step (2): add challenge microorganism: when half hygrometric state resistance bacterium property wet in carrying out dry state resistance bacterium property and testing or do outward is tested, smear by sterile swab and get Challenge and method thing, being inoculated on the outside surface of test sample by challenging room mouth, smearing even; When carrying out half hygrometric state resistance bacterium property test dry in exogenous damp or hygrometric state resistance bacterium property test, in challenge room, add challenge bacterium liquid by challenge room mouth.
Step (3): add TSB substratum: when carrying out half hygrometric state resistance bacterium property test wet in outer doing or hygrometric state resistance bacterium property tests, add TSB substratum by sampler chamber mouth in sampler chamber; When carrying out half hygrometric state resistance bacterium property test dry in dry state resistance bacterium property test or exogenous damp, omit this step.
Step (4): sample challenge: close challenge room mouth and sampler chamber mouth in an adequate manner, guarantee to produce extraneous contamination, puts into incubator and cultivates by testing apparatus.
Step (5): penetrate microorganism checking: when carrying out half hygrometric state resistance bacterium property test dry in dry state resistance bacterium property test or exogenous damp, after cultivation completes, moistening sterile swab squeeze out unnecessary moisture in advance in stroke-physiological saline solution, by all internal surfaces of sampler chamber mouth wiping test sample, then inoculate in the mode on wiping TSA culture medium flat plate surface, postvaccinal TSA culture medium flat plate is put into incubator cultivation and be used for the sampling analysis of sample internal surface.In order to sample fully, the microorganism also in order to be upsampled to by swab is fully inoculated on flat board, and each sample uses three sterile swab wipe samples and inoculates three times, and period repeatedly should rotate swab.
When carrying out half hygrometric state resistance bacterium property test wet in outer doing or hygrometric state resistance bacterium property tests, after having cultivated, observe microorganism growth situation in TSB substratum.
Step (6): challenge microbial activity inspection: when half hygrometric state resistance bacterium property wet in carrying out dry state resistance bacterium property and testing or do outward is tested, moistening sterile swab squeeze out unnecessary moisture in advance in stroke-physiological saline solution, by challenge room mouth wiping test sample all outer surfaces, then inoculate in the mode on wiping TSA culture medium flat plate surface, postvaccinal TSA culture medium flat plate is put into the vigor inspection of incubator cultivation for challenge microorganism.When wiping and inoculation, each sample uses three sterile swab wipe samples and inoculates three times, and period repeatedly should rotate sterile swab.
When carrying out half hygrometric state resistance bacterium property test dry in exogenous damp or hygrometric state resistance bacterium property test, challenge bacterium liquid is dipped by sterile swab, then inoculate in the mode on wiping TSA culture medium flat plate surface, postvaccinal TSA culture medium flat plate is put into the vigor inspection of incubator cultivation for challenge microorganism.
Step (7): result judges: when challenge microorganism has a vigor, test effectively, otherwise invalidate the test; If all without microorganism growth on the substratum of sample internal surface sampling analysis, be judged to and meet the requirement of resistance bacterium property; If have microorganism growth on the substratum of sample internal surface sampling analysis, check whether it is challenge bacterium, if challenge bacterium, be then judged to and do not meet the requirement of resistance bacterium property, if not challenge bacterium, then illustrate there is extraneous contamination, invalidate the test.
Further technical scheme is: when carrying out half hygrometric state resistance bacterium property test or hygrometric state resistance bacterium property test, the incubator of described step (4) adopts shaking culture case and arranges oscillation frequency, to make challenge bacterium liquid fully contact test sample outside surface, or TSB substratum is made fully to contact test sample internal surface.
Further technical scheme is: the determination methods of described step (7) is:
When carrying out dry state resistance bacterium property test, when having challenge bacteria growing on the TSA culture medium flat plate of vigor inspection, test is effective, otherwise invalidate the test; If all without microorganism growth on the TSA culture medium flat plate of sample internal surface sampling analysis, be judged to and meet the requirement of resistance bacterium property; If have microorganism growth on the TSA culture medium flat plate of sample internal surface sampling analysis, check whether it is challenge bacterium, if challenge bacterium, be then judged to and do not meet the requirement of resistance bacterium property, if not challenge bacterium, then illustrate there is extraneous contamination, invalidate the test.
When carrying out half hygrometric state resistance bacterium property test wet in outer doing, when having challenge bacteria growing on the TSA culture medium flat plate of vigor inspection, test effectively, otherwise invalidate the test; If TSB substratum display clarification, be judged to and meet the requirement of resistance bacterium property; If the display of TSB substratum is muddy, checks whether the microorganism in it is challenge bacterium, if challenge bacterium, be then judged to and do not meet the requirement of resistance bacterium property, if not challenge bacterium, then illustrate there is extraneous contamination, invalidate the test.
When carrying out half hygrometric state dry in exogenous damp and hindering the test of bacterium property, when having the growth of challenge bacteria suspension on the TSA culture medium flat plate of vigor inspection, test is effective, otherwise invalidate the test; If all without microorganism growth on the TSA culture medium flat plate of sample internal surface sampling analysis, be judged to and meet the requirement of resistance bacterium property; If have microorganism growth on the TSA culture medium flat plate of sample internal surface sampling analysis, check whether it is challenge bacterium, if challenge bacterium, be then judged to and do not meet the requirement of resistance bacterium property, if not challenge bacterium, then illustrate there is extraneous contamination, invalidate the test.
When carrying out hygrometric state resistance bacterium property test, when having the growth of challenge bacteria suspension on the TSA culture medium flat plate of vigor inspection, test effectively, otherwise invalidate the test; If TSB substratum display clarification, be judged to and meet the requirement of resistance bacterium property; If the display of TSB substratum is muddy, checks whether the microorganism in it is challenge bacterium, if challenge bacterium, be then judged to and do not meet the requirement of resistance bacterium property, if not challenge bacterium, then illustrate there is extraneous contamination, invalidate the test.
The reagent that test method of the present invention is used and material have: serratia marcesens, ATCC8100, or other equivalent bacterial strain; Trypticase soya broth (TSB); Tryptic Soy Agar (TSA); Stroke-physiological saline solution; Sterile swab.
Before testing, challenge microorganism should be prepared, test method challenge microorganism preparation method following (for serratia marcesens) of the present invention:
Prepared by Challenge and method thing: picking serratia marcesens bacterium colony from serratia marcesens work bacterial strain inclined-plane, streak inoculation on TSA flat board, under 25 DEG C of conditions, incubated overnight obtains Challenge and method thing, for dry state resistance bacterium property test and half hygrometric state (outer dry interior wet) resistance bacterium property test.The flat board prepared is before use in 4 DEG C of preservations, and the same day uses.
Challenge bacterium solution preparation: from the preparation of above-mentioned Challenge and method thing obtained with the single colony inoculation of picking serratia marcesens the TSA flat board of Challenge and method thing in TSB substratum, incubated overnight under 25 DEG C of conditions, obtain the challenge bacterium liquid that concentration is about 109cfu/mL, for the test of half hygrometric state (exogenous damp is dry interior) resistance bacterium property.The challenge bacterium liquid prepared is before use in 4 DEG C of preservations, and the same day uses.
The invention has the beneficial effects as follows:
1, provide a kind of testing apparatus, for claiming that the Wound dressing with resistance bacterium performance carries out resistance bacterium benchmark test, the test of the different states such as dry state, half hygrometric state and hygrometric state can be carried out, scientific comprehensive evaluation is carried out to the resistance bacterium performance of Wound dressing;
2, provide a kind of test method, for claiming that the Wound dressing with resistance bacterium performance carries out resistance bacterium performance evaluation, and cover the resistance bacterium performance evaluation of dressing under the different states such as dry state, half hygrometric state, hygrometric state, evaluation result science is comprehensive.
Accompanying drawing explanation
Fig. 1 is the structural representation of testing apparatus of the present invention when carrying out dry state resistance bacterium property test;
Fig. 2 is the structural representation of testing apparatus of the present invention when carrying out half hygrometric state (wet in doing) resistance bacterium property test outward;
Fig. 3 is the structural representation of testing apparatus of the present invention when carrying out half hygrometric state (dry in exogenous damp) resistance bacterium property test;
Fig. 4 is the structural representation of testing apparatus of the present invention when carrying out hygrometric state resistance bacterium property test.
In figure: 1 blocking, 2 testing apparatus main bodys, 21 challenge rooms, 22 sampler chambers, 3 Challenge and method things, 31 serratia marcesens inoculums, 32 challenge bacterium liquid, 4 standing bolts, 5 test samples, 6 sterile swab, 7 supports, 8 bend pipes, 9TSB substratum.
Embodiment
Below in conjunction with Figure of description and specific embodiment, the invention will be further described:
The reagent that the test method of the embodiment of the present invention is used and material have: serratia marcesens, ATCC8100, or other equivalent bacterial strain; Trypticase soya broth (TSB); Tryptic Soy Agar (TSA); Stroke-physiological saline solution; Sterile swab.
Before testing, challenge microorganism should be prepared, the test method challenge microorganism preparation method following (for serratia marcesens) of the embodiment of the present invention:
Prepared by Challenge and method thing: picking serratia marcesens bacterium colony from serratia marcesens work bacterial strain inclined-plane, streak inoculation on TSA flat board, under 25 DEG C of conditions, incubated overnight obtains Challenge and method thing, for dry state resistance bacterium property test and half hygrometric state (outer dry interior wet) resistance bacterium property test.The flat board prepared is before use in 4 DEG C of preservations, and the same day uses.
Challenge bacterium solution preparation: from the preparation of above-mentioned Challenge and method thing obtained with the single colony inoculation of picking serratia marcesens the TSA flat board of Challenge and method thing in TSB substratum, incubated overnight under 25 DEG C of conditions, obtains concentration and is about 109the challenge bacterium liquid of cfu/mL, for the test of half hygrometric state (exogenous damp is dry interior) resistance bacterium property.The challenge bacterium liquid prepared is before use in 4 DEG C of preservations, and the same day uses.
Before test, adopt that to carry out sterilizing through acknowledged sterilizing program to testing apparatus for subsequent use.The test method of following examples all adopts serratia marcesens as challenge bacterium.
Embodiment one:
As shown in Figure 1.A kind of resistance bacterium property testing apparatus, comprise the challenge room 21 for holding Challenge and method thing and the sampler chamber 22 for cleaning sampling, challenge room 21 and sampler chamber 22 are combined and are formed testing apparatus main body 2, be provided with test sample 5 between challenge room 21 and sampler chamber 11, test sample 5 clamps by challenge room 21 and sampler chamber 11.
Testing apparatus main body 2 is arranged on a support 7; Described challenge room 21 and sampler chamber 22 are all in groove-like, the open side challenging room 21 and sampler chamber 22 is provided with the protruding edge of a circle along its edge of opening, and the projection edge of challenge room 21 and the projection of sampler chamber 22 are along mutually docking and being clamped by the standing bolt 4 on described support 7; Challenge room 21 is also provided with challenge room mouth, sampler chamber 22 is also provided with sampler chamber mouth, and challenge room mouth and sampler chamber mouth place are equipped with blocking 1.
Described support 7 comprises bracing frame vertical above the base of level and base, and the top of testing apparatus main body 2 is connected with bracing frame by standing bolt 4 with bottom.
Described challenge room 21 and sampler chamber 22 are all in hemispherical, and challenge room 21 and sampler chamber 22 are arranged symmetrically with thus make testing apparatus main body 2 in spherical, and challenge room mouth is arranged on the center of challenge room 21 sphere, and sampler chamber mouth is arranged on the center of sampler chamber 22 sphere.Challenge room 21 and sampler chamber 22 are set to hemispherical, have the following advantages: one is that circular cross section does not have corner, is convenient to alignment, is also convenient to batch production simultaneously when both combine; Two is that spherical testing apparatus body inner surface is smooth without turning, is convenient to observe and is also convenient to cleaning.
Described challenge room 21, sampler chamber 22, blocking 1 all adopt transparent glass to make.Above-mentioned parts adopt transparent glass to make, and are convenient to operation and the viewing test result of test.
The testing apparatus of the present embodiment is used for carrying out the test of dry state resistance bacterium property.
The test method (test of dry state resistance bacterium property) of the testing apparatus of the present embodiment comprises the following steps:
Step (1): sample clamping: get 3 test samples 5 respectively, makes test sample 5 internal surface towards sampler chamber 22, test sample 5 is clamped between challenge room 21 and sampler chamber 22.Method of clamping should pass through confirmation, guarantee the leakage not occurring to challenge microorganism, by the edge clamping of test sample 5 challenge room 21 and sampler chamber 22 between and clamped by standing bolt 4, thus make test sample 5 vertical be arranged on challenge room 21 and sampler chamber 22 between.For the sample that area is larger, can remainder be cut or be wrapped in around testing apparatus main body 2; For the sample that area is less, the sample starting material that can area be adopted larger are tested, or suitable mode can be adopted to fill lack part for test.
Step (2): add challenge microorganism: smear by sterile swab 6 and get Challenge and method thing 3, and be inoculated on test sample 5 outside surface by challenge room mouth, smear evenly as far as possible.
Step (3): sample challenge: the challenge room mouth of blocking test device and sampler chamber mouth in an adequate manner, guarantees to produce extraneous contamination, testing apparatus is put into 25 DEG C of incubators to 24h.
Step (4): penetrate microorganism checking: after the specified time, moistening sterile swab 6 squeeze out unnecessary moisture in advance in stroke-physiological saline solution, to be tried one's best all internal surfaces of wiping test sample 5 by sampler chamber mouth, then inoculate in the mode of wiping TSA planar surface.In order to sample fully, the microorganism also in order to be upsampled to by swab is fully inoculated on flat board, and each test sample 5 uses three sterile swab wipe samples and inoculates three times, and period should repeatedly rotate sterile swab 6.TSA flat board is placed in 35 DEG C of cultivation 24h and is used for the sampling analysis of sample internal surface.
Step (5): challenge microbial activity inspection: moistening sterile swab 6 squeeze out unnecessary moisture in advance in stroke-physiological saline solution, by challenge room mouth wiping test sample 5 all outer surfaces, then inoculate in the mode on wiping TSA culture medium flat plate surface, each test sample 5 uses three sterile swab wipe samples and inoculates three times, and period should repeatedly rotate sterile swab 6.Postvaccinal TSA culture medium flat plate is placed in 35 DEG C of vigor inspections of cultivating 24h and being used for challenging microorganism.
Step (6): result judges: serratia marcesens should be had on the TSA flat board of vigor inspection to grow, otherwise invalidate the test.If all without microorganism growth on the TSA flat board of sample internal surface sampling analysis, be judged to and meet the requirement of dry state resistance bacterium property.If have microorganism growth on the TSA flat board of sample internal surface sampling analysis, suitable micro-biological process should be adopted to check, and whether it is serratia marcesens.If serratia marcesens, be then judged to and do not meet the requirement of dry state resistance bacterium property; If not serratia marcesens, then illustrate there is extraneous contamination, invalidate the test.
Serratia marcesens can present red colonies at agar surface, and this feature may be used for differentiating whether have serratia marcesens to grow.
Embodiment two:
As shown in Figure 2.The testing apparatus of the present embodiment and the testing apparatus of embodiment one are distinguished and are: described sampler chamber mouth is plugged with a bend pipe 8, and described blocking 1 matches with the end of bend pipe 8, and described bend pipe 8 also adopts transparent glass to make.
The testing apparatus of the present embodiment is used for carrying out half hygrometric state (outer dry interior wet) resistance bacterium property test.
The test method [half hygrometric state (outer dry interior wet) resistance bacterium property test] of the testing apparatus of the present embodiment comprises the following steps:
Step (1): sample clamping: get 3 test samples 5 respectively, makes test sample 5 internal surface towards sampler chamber 22, test sample 5 is clamped between challenge room 21 and sampler chamber 22.Method of clamping should pass through confirmation, guarantee the leakage not occurring to challenge microorganism, by the edge clamping of test sample 5 challenge room 21 and sampler chamber 22 between and clamped by standing bolt 4, thus make test sample 5 vertical be arranged on challenge room 21 and sampler chamber 22 between.For the sample that area is larger, can remainder be cut or be wrapped in around testing apparatus main body 2; For the sample that area is less, the sample starting material that can area be adopted larger are tested, or suitable mode can be adopted to fill lack part for test.
Step (2): add challenge microorganism: smear by sterile swab 6 and get serratia marcesens inoculum 31, and be inoculated on test sample 5 outside surface by challenge room mouth, smear evenly as far as possible.
Step (3): add TSB substratum: add appropriate TSB substratum 9 through sampler chamber mouth in sampler chamber 22 by bend pipe 8.
Step (4): sample challenge: the challenge room mouth of blocking test device and sampler chamber mouth in an adequate manner, guarantee to produce extraneous contamination, testing apparatus is put into 25 DEG C of incubators to 24h, adopt shaking culture case and suitable oscillation frequency is set fully can contacts test sample 5 internal surface to make TSB substratum 9.
Step (5): penetrate microorganism checking: after the specified time, microorganism growth situation in visual inspection TSB substratum 9.
Step (6): challenge microbial activity inspection: moistening sterile swab 6 squeeze out unnecessary moisture in advance in stroke-physiological saline solution, by challenge room mouth wiping test sample 5 all outer surfaces, then inoculate in the mode on wiping TSA culture medium flat plate surface, each test sample 5 uses three sterile swab wipe samples and inoculates three times, and period should repeatedly rotate sterile swab 6.Postvaccinal TSA culture medium flat plate is placed in 35 DEG C of vigor inspections of cultivating 24h and being used for challenging microorganism.
Step (7): result judges: serratia marcesens should be had on the TSA flat board of vigor inspection to grow, otherwise invalidate the test.If TSB substratum display clarification, be judged to and meet half hygrometric state (outer dry interior wet) resistance bacterium property requirement.If the display of TSB substratum is muddy, suitable micro-biological process should be adopted to check, and whether it is serratia marcesens.If serratia marcesens, be then judged to and do not meet half hygrometric state (outer dry interior wet) resistance bacterium property requirement; If not serratia marcesens, then illustrate there is extraneous contamination, invalidate the test.
If necessary, the TSB substratum of aseptic TSB substratum as negative control (display clarification) and inoculation serratia marcesens can be set as positive control (display is muddy), clarify to avoid visual inspection or muddy possible operative error.
Embodiment three:
As shown in Figure 3.The testing apparatus of the present embodiment and the testing apparatus of embodiment one are distinguished and are: described challenge room mouth is plugged with a bend pipe 8, and described blocking 1 matches with the end of bend pipe 8, and described bend pipe 8 also adopts transparent glass to make.
The testing apparatus of the present embodiment is used for carrying out the test of half hygrometric state (dry in exogenous damp) resistance bacterium property.
The test method [test of half hygrometric state (dry in exogenous damp) resistance bacterium property] of the testing apparatus of the present embodiment comprises the following steps:
Step (1): sample clamping: get 3 test samples 5 respectively, makes test sample 5 internal surface towards sampler chamber 22, test sample 5 is clamped between challenge room 21 and sampler chamber 22.Method of clamping should pass through confirmation, guarantee the leakage not occurring to challenge microorganism, by the edge clamping of test sample 5 challenge room 21 and sampler chamber 22 between and clamped by standing bolt 4, thus make test sample 5 vertical be arranged on challenge room 21 and sampler chamber 22 between.For the sample that area is larger, can remainder be cut or be wrapped in around testing apparatus main body 2; For the sample that area is less, the sample starting material that can area be adopted larger are tested, or suitable mode can be adopted to fill lack part for test.
Step (2): add challenge microorganism: add appropriate challenge bacterium liquid 32 through challenge room mouth in challenge room 21 by bend pipe 8.
Step (3): sample challenge: the challenge room mouth of blocking test device and sampler chamber mouth in an adequate manner, guarantee to produce extraneous contamination, testing apparatus is put into 25 DEG C of incubators to 24h, adopt shaking culture case and suitable oscillation frequency is set fully can contacts test sample 5 outside surface to make challenge bacterium liquid 32.
Step (4): penetrate microorganism checking: after the specified time, moistening sterile swab 6 squeeze out unnecessary moisture in advance in stroke-physiological saline solution, to be tried one's best all internal surfaces of wiping test sample 5 by sampler chamber mouth, then inoculate in the mode of wiping TSA planar surface.Each test sample 5 uses three sterile swab wipe samples and inoculates three times, and period should repeatedly rotate sterile swab 6.TSA flat board is placed in 35 DEG C of cultivation 24h and is used for the sampling analysis of sample internal surface.
Step (5): challenge microbial activity inspection: dip challenge bacterium liquid 32 wiping inoculation TSA by sterile swab 6 dull and stereotyped, postvaccinal TSA culture medium flat plate is placed in 35 DEG C of vigor inspections of cultivating 24h and being used for challenging microorganism.
Step (6): result judges: serratia marcesens suspension should be had on the TSA flat board of vigor inspection to grow, otherwise invalidate the test.If all without microorganism growth on the TSA flat board of sample internal surface sampling analysis, be judged to and meet the requirement of half hygrometric state (dry in exogenous damp) resistance bacterium property.If have microorganism growth on the TSA flat board of sample internal surface sampling analysis, suitable micro-biological process should be adopted to check, and whether it is serratia marcesens.If serratia marcesens, be then judged to and do not meet the requirement of half hygrometric state (dry in exogenous damp) resistance bacterium property; If not serratia marcesens, then illustrate there is extraneous contamination, invalidate the test.
Embodiment four:
As shown in Figure 4.The testing apparatus of the present embodiment and the testing apparatus of embodiment one are distinguished and are: described challenge room mouth and sampler chamber mouth are all plugged with a bend pipe 8, and described blocking 1 matches with the end of bend pipe 8, and described bend pipe 8 also adopts transparent glass to make.
The testing apparatus of the present embodiment is used for carrying out the test of hygrometric state resistance bacterium property.
The test method (test of hygrometric state resistance bacterium property) of the testing apparatus of the present embodiment comprises the following steps:
Step (1): sample clamping: get 3 test samples 5 respectively, makes test sample 5 internal surface towards sampler chamber 22, test sample 5 is clamped between challenge room 21 and sampler chamber 22.Method of clamping should pass through confirmation, guarantee the leakage not occurring to challenge microorganism, by the edge clamping of test sample 5 challenge room 21 and sampler chamber 22 between and clamped by standing bolt 4, thus make test sample 5 vertical be arranged on challenge room 21 and sampler chamber 22 between.For the sample that area is larger, can remainder be cut or be wrapped in around testing apparatus main body 2; For the sample that area is less, the sample starting material that can area be adopted larger are tested, or suitable mode can be adopted to fill lack part for test.
Step (2): add challenge microorganism: add appropriate challenge bacterium liquid 32 through challenge room mouth in challenge room 21 by bend pipe 8.
Step (3): add TSB substratum: add appropriate TSB substratum 9 through sampler chamber mouth in sampler chamber 22 by bend pipe 8.
Step (4): sample challenge: the challenge room mouth of blocking test device and sampler chamber mouth in an adequate manner, guarantee to produce extraneous contamination, testing apparatus is put into 25 DEG C of incubators to 24h, adopt shaking culture case and suitable oscillation frequency is set and fully can contact test sample 5 outside surface to make challenge bacterium liquid 32, make TSB substratum 9 fully can contact test sample 5 internal surface simultaneously.
Step (5): penetrate microorganism checking: after the specified time, microorganism growth situation in visual inspection TSB substratum 9.
Step (6): challenge microbial activity inspection: dip challenge bacterium liquid 32 wiping inoculation TSA by sterile swab 6 dull and stereotyped, postvaccinal TSA culture medium flat plate is placed in 35 DEG C of vigor inspections of cultivating 24h and being used for challenging microorganism.
Step (7) result judges: serratia marcesens suspension should be had on the TSA flat board of vigor inspection to grow, otherwise invalidate the test.If TSB substratum display clarification, be judged to and meet the requirement of hygrometric state resistance bacterium property.If the display of TSB substratum is muddy, suitable micro-biological process should be adopted to check, and whether it is serratia marcesens.If serratia marcesens, be then judged to and do not meet the requirement of hygrometric state resistance bacterium property; If not serratia marcesens, then illustrate there is extraneous contamination, invalidate the test.
Test method of the present invention needs to operate microorganism, should be tested by the personnel through training in biocontainment laboratory.
The foregoing is only preferred embodiment of the present invention; be not whole embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all; any amendment of doing, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Except technical characteristic described in specification sheets, all the other technical characteristics are those skilled in the art's known technology, and be outstanding innovative characteristics of the present invention, above-mentioned technical characteristic does not repeat them here.