CN201793571U - Isolating device detection system used for biological detection of efficient particle filter - Google Patents
Isolating device detection system used for biological detection of efficient particle filter Download PDFInfo
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- CN201793571U CN201793571U CN2010205186593U CN201020518659U CN201793571U CN 201793571 U CN201793571 U CN 201793571U CN 2010205186593 U CN2010205186593 U CN 2010205186593U CN 201020518659 U CN201020518659 U CN 201020518659U CN 201793571 U CN201793571 U CN 201793571U
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- particle filter
- highly effective
- effective particle
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Abstract
The utility model relates to an isolating device detection system used for biological detection of an efficient particle filter, which is characterized in that the isolating device detection system comprises a sealed isolation bin, wherein the top of the isolation bin is provided with an air inlet connected with an air inlet pipe, and an air outlet connected with an air outlet pipe, the air inlet and the air outlet are internally and respectively provided with the efficient particle filter, and a microbial aerosol generator and an air microbial sampler are arranged at the front end of the efficient particle filter at the air outlet; an isolating room is externally provided with one to four air microbial samplers, and the air inlet end of each sampler is communicated into the air outlet pipe above the rear end of the efficient particle filter at the air outlet by a sampling pipe; and each sampler is internally provided with a microbial culture plate, and the outlets of samplers are respectively connected with an aspirator pump by pipelines. The isolating device detection system can be widely applied in an injected animal feeding negative pressure isolating device, is safe and reliable to operate, true and accurate for detection effect, and does not generate secondary pollution to the environment.
Description
Technical field
The utility model relates to a kind of Biological Detection device, particularly about the isolation device detection system that highly effective particle filter biological detects that is used in a kind of animal rearing negative pressure disrupter.
Background technology
In the medical microbiology research and experiment; often to carry out the zoogenetic infection experimental study; infection animal must be raised at infection animal and be raised in the disrupter; it is the prerequisite Biosafety shield of infection animal safe feeding in the pathogenic micro-organism laboratory that infection animal is raised disrupter; it is the one-level protective barrier of three grades and level Four biocontainment laboratory; infection animal is raised the ventilation and negative pressure ventilation filtering system of disrupter to the aerocolloidal filtration, purification effect of pathogenic micro-organism, is directly connected to the protection to the inside and outside environment of biocontainment laboratory.
At present, to the detection of highly effective particle filter in the ventilation and negative pressure ventilation system of infection animal raising disrupter, all be to adopt abiotic aerocolloidal method both at home and abroad, promptly adopt physics methods such as particle scanning, photo densitometry to detect evaluation.During detection, the general monodisperse aerosol (0.3 μ m or 0.5 μ m) that uses artificial generation dioctyl phthalate (DOP) (DOP), poly-alpha olefins (PAO), polyoxyethylene glycol etc., the detection of leaking.Because bioaerosol particle and abiotic aerosol particles exist at aspects such as moiety, structure, physical propertys than big-difference, particularly the microbial aerosol that produces of laboratory be polydisperse, particle diameter is little, institute is electrically charged also different because of microbe species.Therefore adopt existing method the negative pressure infection animal to be raised the detection evaluation result of highly effective particle filter in the exhaust system of disrupter, often be not inconsistent with the actual filtration effect of highly effective particle filter to microbial aerosol.Simultaneously conventional detection only to highly effective particle filter detections of lining by line scan, can't be leaked the structure of whole highly effective particle filter, damage leakage, material structure leakage make systematic detection.The physics detection method also exists the complicacy and the demanding characteristics of instrument of execute-in-place in addition.
Summary of the invention
At the problems referred to above, the purpose of this utility model provides a kind of isolation device detection system that highly effective particle filter biological detects that is used for, and it can be applied in infection animal and raise in the negative pressure disrupter, and operational safety is reliable, detect the effect true and accurate, can not produce secondary pollution environment.
For achieving the above object, the utility model is taked following technical scheme: a kind of isolation device detection system that is used for the highly effective particle filter biological detection, it is characterized in that: it comprises the isolation cabin of a sealing, top at described isolation cabin, one air outlet that connects blast tube is connected exhaust duct with one exhaust outlet is set, one highly effective particle filter is set respectively in described air outlet and exhaust outlet, and the front end of described exhaust outlet place highly effective particle filter is provided with a microbial aerosol producer and an air microorganism sampler; The outside is provided with 1～4 air microorganism sampler between described isolation, and the inlet end of each sampling thief feeds in the exhaust duct of top, highly effective particle filter rear end, described exhaust outlet place by a sampling tube respectively; Be provided with the microorganism culturing ware in each described sampling thief, and the outlet of each described sampling thief connects off-gas pump by a pipeline respectively; Air enters from described air outlet, discharges from described exhaust duct through the highly effective particle filter filtration back at described exhaust outlet place.
The sampling thief of described highly effective particle filter front end is six grades of air microorganism samplers of Anderson.
The sampling thief of described highly effective particle filter rear end is three Anderson secondary air microorganism samplers, and described sampling thief is arranged on place, exhaust outlet axis, left side, axis and the right side, axis at the described highly effective particle filter exhaust outlet 40～60cm of distance place.
The utility model is owing to take above technical scheme, it has the following advantages: 1, the utility model can adopt the selected simulation indicator microoraganism of using of microbial aerosol, be bacterium and the virus (comprising phage) that does not have in the air, and this bacterium all is safe from harm to crowd, animal, environment etc. with virus, therefore, testing process is safe and reliable, the detected result true and accurate, and specificity is very high.2, the utility model can select serratia marcescens to substitute malignant bacteria, also can select the phage of serratia marcescens to substitute Causative virus, collect the aerosol particles of highly effective particle filter front and back by the nutrient agar that is arranged in the collector simultaneously, and by carrying out serratia marcescens bacterium colony and phage plaque counting after the bacterium that falls is cultivated, and then by calculating the actual filtration effect of highly effective particle filter, realize highly effective particle filter biological detection method, remedied the defective of existing physics detection method.3, systematicness of the present utility model is strong, and whether not only can detect highly effective particle filter has leakage, can also detect the framework of installing between highly effective particle filter and the negative pressure infection animal raising disrupter and whether reach airtight sealing.5, the utility model practical simplicity not only, real result is accurate, and the process that the utility model is raised at infection animal, and is more safe and reliable.The utility model can be widely used in the highly effective particle filter biological testing process under the various closed negative pressure conditions.
Description of drawings
Fig. 1 is a structural representation of the present utility model
Embodiment
Below in conjunction with drawings and Examples, the utility model is described in detail.
As shown in Figure 1, the utility model comprises the isolation cabin 1 of a sealing, top at isolation cabin 1, one air outlet 2 that connects blast tube is connected exhaust duct with one exhaust outlet 3 is set, one highly effective particle filter (not shown) is set in air outlet 2 and exhaust outlet 3, and the utility model mainly is the environmental pollution that the control air draft may cause.The utility model is at the isolation cabin 1 inside and outside air microorganism sampler 4,5 that is respectively arranged with, in order to make the detection better effects if, 1～4 sampling thief 5 can be set outside isolation cabin 1, the inlet end of each sampling thief 5 is connected in the exhaust duct of exhaust outlet 3 tops by a sampling tube 6 respectively, outlet at sampling thief 4,5 connects an off-gas pump (not shown) by a pipeline respectively, and an off-gas pump also can two pipelines be connected in parallel.In isolation cabin 1 outside or a microbial aerosol producer 7 is set in isolation cabin 1, the inlet end of microbial aerosol producer 7 is identical with ordinary skill, connects a under meter, pressure warning unit, oil-filtering apparatus and an air compressor (not shown) successively by pipeline.If the space in the isolation cabin 1 is bigger, microbial aerosol producer 7 can be arranged in the isolation cabin 1; If microbial aerosol producer 7 is arranged on isolation cabin 1 outside, then be connected to air outlet 2 places in the isolation cabin 1 by a pipeline 8 (for example rubber hose) in its outlet side, the air flow line that the outlet of pipeline and blast tube are sent into is consistent substantially.
The artificial alternative abiotic particle of simulation indicator microoraganism that takes place of microbial aerosol producer 7 usefulness in the utility model.Wherein simulate indicator microoraganism and comprise two kinds, a kind of is bacterium, and with what do not have in the natural air, to the bacterium of people, animal, the harm of environment lifeless matter, another kind is a virus, with what do not have in the natural air, to the virus of people, animal, the harm of environment lifeless matter.Bacterium of the present utility model is used serratia marcescens, also can use other non-pathogenic bacteria; Virus of the present utility model is used the phage of serratia marcescens, also can use other nonpathogenic virus, phage.The utility model is carrying out highly effective particle filter in the process of Biological Detection, be manually to contain the simulation indicator microoraganism aerosol of bacterium respectively and contain viral simulation indicator microoraganism aerosol by microbial aerosol producer 7, and collect by being arranged on isolation cabin 1 inside and outside sampling thief 4,5 respectively, again the bacterium or the virus of collecting are carried out biology cultivation and counting, by calculating the proportionlity that highly effective particle filter filters forward and backward count results, obtain the true filtration efficiency of the corresponding microorganism of highly effective particle filter then.
Below by specific embodiment, the utility model is further described.
The material that the utility model uses in carrying out the Biological Detection process:
1, simulation indicator microoraganism:
The bacterium of present embodiment is adopted serratia marcescens (Serratia Marcescens), and the serratia marcescens bacterium colony is a rose, and dimpling is smooth moistening, neat in edge.
Virus of the present utility model adopts serratia marcescens phage SM701 suspension.
The artificial particle diameter scope that the simulation microbial aerosol takes place is between 0.5～3 μ m, and wherein the microbial aerosol population of 1 μ m accounts for 75%;
2, reagent
(1) phosphate buffered saline buffer (PBS, 0.03mol/L, pH7.2)
Get disodium hydrogen phosphate,anhydrous 2.83 grams, potassium primary phosphate 1.36 grams add distilled water to 1000ml, regulate pH to 7.2～7.4, and sterilization 20min is standby under 121 ℃ of high pressure steam.
(2) plain agar substratum
Composition: peptone 10 gram extractum carnis powder 3 gram NaCl 5 gram agar powders 15 grams
Add the fusion of 1000ml distilled water or deionized water Hybrid Heating, regulate pH to 7.2～7.4, sterilization 20min is standby under 121 ℃ of high pressure steam.
(3) broth medium
Composition: extractum carnis 3 gram peptones 10 gram sodium-chlor 5 grams
Add water to 1000ml, regulate pH to 7.2～7.4, sterilization 20min is standby under 121 ℃ of high pressure steam.
(4) LB substratum (Luria-Bertani substratum)
Composition: Tryptones 10 gram yeast extracts 5 gram NaCl 10 grams
Add the fusion of 1000ml distilled water or deionized water Hybrid Heating, regulate pH to 7.2～7.4, sterilization 20min is standby under 121 ℃ of high pressure steam.
3, serratia marcescens suspension preparation
(1) under aseptic condition, serratia marcescens (Serratia Marcescens) bacterial classification streak inoculation is arrived plain agar substratum plate (on 90 * 15mm);
(2) cultivate 24 ± 2h down at 30 ± 0.5 ℃;
(3) under aseptic condition from characteristic bacterium colony (rose, dimpling, smooth moistening, the neat in edge) surface picking one transfering loop bacterium transfer in the test tube that 5ml meat soup liquid nutrient medium is housed;
(4) under 37 ± 0.5 ℃, shaking table (160rpm) is cultivated 24 ± 2h;
(5) preserve down at 4 ℃, the storage life is 15 days, determines bacterial concentration with the plate dilution method of counting before using, Zhi Bei bacterium liquid as stated above, and its mean concns should be 1 * 10
10～4 * 10
10/ ml;
(6) with the PBS diluent bacterium stoste can be used by after the suitable proportion dilution.
4, the preparation and the cultivation of serratia marcescens phage SM701 suspension
(1) preparation of host bacterium bacterium liquid
The serratia marcescens that to grow on the plain agar substratum is inoculated in the meat soup liquid nutrient medium, and under 37 ℃, 150rpm shaking culture 8h preserves standby down at 4 ℃.
(2) propagation of serratia marcescens phage SM701
Serratia marcescens phagocytosis body fluid 0.1ml and host bacterium liquid 0.2ml are added in the 5ml LB liquid nutrient medium, room temperature is placed 1h, behind 37 ℃ of following 50rpm shaking culture 3.5h, and the centrifugal 10min of whizzer 10000rpm, with 0.22 μ m membrane filtration, gained amplification liquid is tired and is checked and should be 10 with supernatant liquor
9～10
10Pfu/ml preserves down at 4 ℃.As the deficiency of tiring, can repeat propagation.
(3) phage is cultivated counting:
Double-deck Plating: lower floor puts into sampling thief 4,5 and carries out taking out after the phage sampling with the LB substratum 7～9ml shop ware that contains 1.5% agar, under aseptic condition, adds in upper strata semisolid medium 4～6ml of 50 ℃ and contains 0.3ml 10
9The serratia marcescens of pfu/ml is placed on the table top then and shakes up, and makes the upper strata substratum be paved with flat board, after waiting to solidify, cultivates 12～24h down at 30～36 ℃, the counting plaque.
Adopt the utility model to carry out biological detection method, may further comprise the steps:
1, background detects
2, should satisfy following condition to the selection of microbial aerosol producer 7:
(1) gas blowout, downdraught type producer;
(2) the aerosolized serratia marcescens liquid 〉=0.2ml of per minute under the nominal operation flow;
(3) the following sub-ratio of bioplast 〉=80% of particle diameter 2 μ m in the serratia marcescens aerosol of Shi Fanging;
(4) release rate of microbial aerosol is 30 ± 3m/min.
3, following method is adopted in 7 calibrations to the microbial aerosol producer:
(1) area (m of the fog nozzle of measurement microbial aerosol producer
2), when calculating the speed ejection aerosol with 0.5m/s, the airshed (m of aerosol dispenser
3/ s);
(2) the husky Lei Shi suspension of the cement bacterium of adding certain volume in microbial aerosol producer 7, suspension bacterium colony concentration is calculated with plate dilution method;
(3) open microbial aerosol producer 7 and sampling thief 4, the control airshed, making aerosol ejection speed is 0.5m/s, to the damage rate of microorganism less than 15%.Shunt phegma after the aerosol dispenser work, accurately calculate its bacterium colony concentration with plate dilution method after the measurement volumes, sampling 20min.Accurate measurement in sampling back and record residue bacteria liquid are long-pending;
(4) result calculates
Aerosol generating capacity=(bacteria liquid amasss-the phegma volume behind bacterium liquid original volume-spraying 20min)/20;
Aerosol ejection speed=air flow rate (m
3/ s)/fog nozzle area (m
2);
The sub-ratio of the following bioplast of particle diameter 2 μ m in the aerosol=4,5 grades of plate colony numbers of Andersen formula sampling thief/plate colony number sums at different levels
The rate of recovery of serratia marcescens (PBS diluent)=phegma bacterium colony concentration/bacterium liquid original content.
When present embodiment detects, use the PBS diluted to (1～8) * 10 the serratia marcescens suspension
7/ ml, ice bath is placed, and is connected on the fluid inlet of microbial aerosol producer 7 in the utility model device, and the particle diameter scope that the mimic microbial aerosol takes place is between 0.5～3 μ m, and wherein the microbial aerosol population of 1 μ m accounts for 75%.
4, the acquisition testing of exhaust outlet analog sample
Place, exhaust outlet 3 axis at 40～60cm place, left side, axis and right side, axis above distance isolation cabin 1 exhaust outlet 3, one sampling tube 6 is set respectively, the outlet side that every sampling tube connects 5, three sampling thiefs 5 of an Anderson secondary air microorganism sampler respectively off-gas pump that can be connected in parallel.
Keep that isolation cabin 1 is in normally, under the steady operational status, simulation indicator microoraganism aerosol 10min takes place isolation cabin 1 in after, begin detection and sample.Sampling time is 10min, and sampling thief 5 flows are 28.3L/min, and every kind (comprising bacterium and virus) detected and repeat 3 times.
5, test sample culture assays
After background gathered sample, analog detection and gather sample and place 30 ± 0.5 ℃ of incubators to cultivate 24 ± 2h, carry out serratia marcescens bacterium colony and phage plaque counting.
6, the calculating of detected result:
(1) bacterium colony concentration is with CFU/m
3Expression, the indication bacteria concentration should be 0CFU/m in the background
3
(2) averaging analog indicator aerosol load should be greater than 10 in the isolation cabin 1
4CFU/m
3Whether indicator aerosol load and highly effective particle filter filtered back indicator aerosol load and calculate before exhaust outlet 3 place's highly effective particle filters should filter according to highly effective particle filter the aerocolloidal filterability of indicator, qualifiedly should judge according to relevant national standard.
The utility model only describes with the foregoing description; the structure of each parts, the position is set and connects and all can change to some extent; on the basis of technical solutions of the utility model; all improvement and equivalents of individual component being carried out according to the utility model principle all should not got rid of outside protection domain of the present utility model.
Claims (3)
1. one kind is used for the isolation device detection system that highly effective particle filter biological detects, it is characterized in that: it comprises the isolation cabin of a sealing, top at described isolation cabin, one air outlet that connects blast tube is connected exhaust duct with one exhaust outlet is set, one highly effective particle filter is set respectively in described air outlet and exhaust outlet, and the front end of described exhaust outlet place highly effective particle filter is provided with a microbial aerosol producer and an air microorganism sampler; The outside is provided with 1～4 air microorganism sampler between described isolation, and the inlet end of each sampling thief feeds in the exhaust duct of top, highly effective particle filter rear end, described exhaust outlet place by a sampling tube respectively; Be provided with the microorganism culturing ware in each described sampling thief, and the outlet of each described sampling thief connects off-gas pump by a pipeline respectively; Air enters from described air outlet, discharges from described exhaust duct through the highly effective particle filter filtration back at described exhaust outlet place.
2. a kind of isolation device detection system that highly effective particle filter biological detects that is used for as claimed in claim 1, it is characterized in that: the sampling thief of described highly effective particle filter front end is six grades of air microorganism samplers of Anderson.
3. a kind of isolation device detection system that highly effective particle filter biological detects that is used for as claimed in claim 1 or 2, it is characterized in that: the sampling thief of described highly effective particle filter rear end is three Anderson secondary air microorganism samplers, and described sampling thief is arranged on place, exhaust outlet axis, left side, axis and the right side, axis at the described highly effective particle filter exhaust outlet 40～60cm of distance place.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010205186593U CN201793571U (en) | 2010-09-03 | 2010-09-03 | Isolating device detection system used for biological detection of efficient particle filter |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010205186593U CN201793571U (en) | 2010-09-03 | 2010-09-03 | Isolating device detection system used for biological detection of efficient particle filter |
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| Publication Number | Publication Date |
|---|---|
| CN201793571U true CN201793571U (en) | 2011-04-13 |
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| CN2010205186593U Expired - Lifetime CN201793571U (en) | 2010-09-03 | 2010-09-03 | Isolating device detection system used for biological detection of efficient particle filter |
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2010
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20110413 |