JP2001523549A - Gelatin membrane filter and method for producing the same - Google Patents
Gelatin membrane filter and method for producing the sameInfo
- Publication number
- JP2001523549A JP2001523549A JP2000520893A JP2000520893A JP2001523549A JP 2001523549 A JP2001523549 A JP 2001523549A JP 2000520893 A JP2000520893 A JP 2000520893A JP 2000520893 A JP2000520893 A JP 2000520893A JP 2001523549 A JP2001523549 A JP 2001523549A
- Authority
- JP
- Japan
- Prior art keywords
- gelatin
- membrane filter
- membrane
- gelatin membrane
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000012528 membrane Substances 0.000 title claims abstract description 118
- 108010010803 Gelatin Proteins 0.000 title claims abstract description 96
- 229920000159 gelatin Polymers 0.000 title claims abstract description 96
- 239000008273 gelatin Substances 0.000 title claims abstract description 96
- 235000019322 gelatine Nutrition 0.000 title claims abstract description 96
- 235000011852 gelatine desserts Nutrition 0.000 title claims abstract description 96
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 15
- 244000005700 microbiome Species 0.000 claims abstract description 33
- 230000015271 coagulation Effects 0.000 claims abstract description 13
- 238000005345 coagulation Methods 0.000 claims abstract description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- 239000000126 substance Substances 0.000 claims description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
- 230000001681 protective effect Effects 0.000 claims description 11
- KYWVDGFGRYJLPE-UHFFFAOYSA-N trimethylazanium;acetate Chemical compound CN(C)C.CC(O)=O KYWVDGFGRYJLPE-UHFFFAOYSA-N 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 230000003204 osmotic effect Effects 0.000 claims description 8
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 claims description 7
- KXKVLQRXCPHEJC-UHFFFAOYSA-N acetic acid trimethyl ester Natural products COC(C)=O KXKVLQRXCPHEJC-UHFFFAOYSA-N 0.000 claims description 7
- 239000011230 binding agent Substances 0.000 claims description 7
- 238000012958 reprocessing Methods 0.000 claims description 4
- 239000010409 thin film Substances 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 3
- 239000010408 film Substances 0.000 claims description 3
- 239000000758 substrate Substances 0.000 claims description 3
- 241000894006 Bacteria Species 0.000 abstract description 14
- 230000001580 bacterial effect Effects 0.000 abstract description 8
- 239000001963 growth medium Substances 0.000 abstract description 3
- 241000233866 Fungi Species 0.000 abstract description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 abstract description 2
- 241000700605 Viruses Species 0.000 abstract description 2
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 235000013305 food Nutrition 0.000 abstract description 2
- 239000002699 waste material Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 18
- 239000000725 suspension Substances 0.000 description 13
- 238000012360 testing method Methods 0.000 description 11
- 239000007789 gas Substances 0.000 description 8
- 229920001817 Agar Polymers 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 238000012937 correction Methods 0.000 description 6
- 244000063299 Bacillus subtilis Species 0.000 description 5
- 235000014469 Bacillus subtilis Nutrition 0.000 description 5
- 239000011159 matrix material Substances 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 4
- 239000000443 aerosol Substances 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 3
- 229920002451 polyvinyl alcohol Polymers 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- KWIUHFFTVRNATP-UHFFFAOYSA-N Betaine Natural products C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 2
- 241000191070 Escherichia coli ATCC 8739 Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- KWIUHFFTVRNATP-UHFFFAOYSA-O N,N,N-trimethylglycinium Chemical compound C[N+](C)(C)CC(O)=O KWIUHFFTVRNATP-UHFFFAOYSA-O 0.000 description 2
- 239000004372 Polyvinyl alcohol Substances 0.000 description 2
- 239000003463 adsorbent Substances 0.000 description 2
- 229960003237 betaine Drugs 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000002906 microbiologic effect Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 231100000614 poison Toxicity 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 239000003440 toxic substance Substances 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- FJWGYAHXMCUOOM-QHOUIDNNSA-N [(2s,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6s)-4,5-dinitrooxy-2-(nitrooxymethyl)-6-[(2r,3r,4s,5r,6s)-4,5,6-trinitrooxy-2-(nitrooxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-3,5-dinitrooxy-6-(nitrooxymethyl)oxan-4-yl] nitrate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O)O[C@H]1[C@@H]([C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@@H](CO[N+]([O-])=O)O1)O[N+]([O-])=O)CO[N+](=O)[O-])[C@@H]1[C@@H](CO[N+]([O-])=O)O[C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O FJWGYAHXMCUOOM-QHOUIDNNSA-N 0.000 description 1
- 238000012387 aerosolization Methods 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- 230000005757 colony formation Effects 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000013100 final test Methods 0.000 description 1
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- 229940049906 glutamate Drugs 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 229960002743 glutamine Drugs 0.000 description 1
- 239000012456 homogeneous solution Substances 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 229960003646 lysine Drugs 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000006199 nebulizer Substances 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 230000001473 noxious effect Effects 0.000 description 1
- 230000008723 osmotic stress Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 229960002429 proline Drugs 0.000 description 1
- 238000000275 quality assurance Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 239000008174 sterile solution Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 229940074410 trehalose Drugs 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 238000009827 uniform distribution Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/22—Devices for withdrawing samples in the gaseous state
- G01N1/2202—Devices for withdrawing samples in the gaseous state involving separation of sample components during sampling
- G01N1/2205—Devices for withdrawing samples in the gaseous state involving separation of sample components during sampling with filters
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
- B01D69/14—Dynamic membranes
- B01D69/141—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes
- B01D69/1411—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes containing dispersed material in a continuous matrix
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
- B01D71/06—Organic material
- B01D71/74—Natural macromolecular material or derivatives thereof
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/22—Devices for withdrawing samples in the gaseous state
- G01N1/2202—Devices for withdrawing samples in the gaseous state involving separation of sample components during sampling
- G01N2001/222—Other features
- G01N2001/2223—Other features aerosol sampling devices
Landscapes
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dispersion Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Sampling And Sample Adjustment (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
(57)【要約】 この発明は増大した数の生存可能の微生物を維持しながら気体から微生物を収集するためのゼラチン膜フィルタならびにその製造方法に関する。これはゼラチン膜フィルタが膜吸引溶液かまたは第１の凝固槽かそのいずれかに添加された浸透保護物質を含有することにより達成される。この発明によるゼラチン膜フィルタは、培地の細菌数負担を同定するために、製薬工業、バイオテクノロジー工業および食品工業、環境保護、廃棄物処理・再利用および医療機器に使用可能である。このゼラチン膜フィルタはチャンバ内でのそれらの濃度を同定できるため、たとえば細菌類、ウィルス類、酵母類および菌類を収集するための空気細菌収集器と組み合せて利用される。 (57) [Summary] The present invention relates to a gelatin membrane filter for collecting microorganisms from a gas while maintaining an increased number of viable microorganisms, and a method of making the same. This is achieved by the fact that the gelatin membrane filter contains a osmoprotectant added to either the membrane suction solution or the first coagulation bath. The gelatin membrane filter according to the present invention can be used in the pharmaceutical, biotechnology and food industries, environmental protection, waste treatment / reuse and medical equipment to identify the bacterial burden on the culture medium. The gelatin membrane filters can be used in combination with an air bacteria collector to collect, for example, bacteria, viruses, yeasts, and fungi because their concentration in the chamber can be identified.
Description
【０００１】 （技術分野） この発明は生存可能な微生物を数多く維持しながら気体から微生物を収集する
ためのゼラチン膜フィルタならびにその製造方法に関する。TECHNICAL FIELD The present invention relates to a gelatin membrane filter for collecting microorganisms from a gas while maintaining a large number of viable microorganisms, and a method for producing the same.
【０００２】 （背景技術） この発明によるゼラチン膜フィルタは気体状の培地から、特に空気から微生物
を収集するために製薬工業、バイオテクノロジー工業および食品工業、環境保護
、廃棄物処理・再利用におよび培地の細菌数負担を同定するための医療機器に使
用可能である。このゼラチン膜フィルタは室内でそれらの濃度を同定できるよう
にするため、たとえば細菌類、ウィルス類、酵母類および菌類を収集するための
空気細菌収集器と組み合せて利用される。このような監視は、たとえば室内空気
中の微生物濃度が高すぎることによる損傷から人員および製品を守るための措置
を適時に導入するための前提条件である。BACKGROUND OF THE INVENTION Gelatin membrane filters according to the present invention are used in the pharmaceutical, biotechnology and food industries, environmental protection, waste treatment and recycling for collecting microorganisms from gaseous media, especially from air. It can be used in medical devices to identify the bacterial burden on a medium. This gelatin membrane filter is used in combination with an air bacteria collector for collecting bacteria, viruses, yeasts and fungi, for example, so that their concentrations can be identified indoors. Such monitoring is a prerequisite for the timely introduction of measures to protect personnel and products from damage, for example due to too high a concentration of microorganisms in the room air.
【０００３】 空気調節された空間、クリーンルームおよび絶縁装置等のような空気の性状に
対し特別の要件を有する特定の空間内の空気が規則的にその細菌濃度を検査され
る。これは当然低濃度の微生物を含む一般に濾過された空気なので、確言力のあ
る結果で細菌を十分に収集するため普通は大容量を検査する必要がある。そのた
め試料空気が適切なフィルタで濾過される。この目的に用いられるフィルタとし
て、たとえばドイツ連邦共和国特許公報第１１７３６４０号明細書に記述されて
いるように、主として硝酸セルロース、酢酸セルロースおよびゼラチンを基材と
する微細濾過領域の小孔サイズを有する滅菌された膜フィルタが使用される。特
に適しているものは抑制された微生物が湿して増殖能力を保持されるべきゼラチ
ン膜フィルタである。試料抽出後にゼラチン膜フィルタは寒天培養基で熟成する
ことができ、個別的に収集された細菌凝集から微生物コロニーが増殖し（ゼラチ
ン膜フィルタが溶解されて消失し、微生物コロニーが直接寒天上で計数すること
ができる）、あるいはペプトン水または生理食塩水のような滅菌溶液の中に溶解
され、その結果部分量を種々の培養基で熟成することができる。[0003] Air in specific spaces that have special requirements for the nature of the air, such as air-conditioned spaces, clean rooms, and insulation, are regularly tested for their bacterial concentration. Since this is, of course, generally filtered air containing low concentrations of microorganisms, large volumes usually need to be tested for sufficient collection of bacteria with robust results. Therefore, the sample air is filtered by a suitable filter. Filters used for this purpose include, for example, sterile filters having a small pore size in the microfiltration area based on cellulose nitrate, cellulose acetate and gelatin, as described in DE-A-1 173 640. The used membrane filter is used. Especially suitable are gelatin membrane filters in which the suppressed microorganisms are to be kept moist and capable of growing. After sample extraction, the gelatin membrane filter can be aged in agar medium, and microbial colonies grow from individually collected bacterial clumps (the gelatin membrane filter dissolves and disappears, and the microbial colonies are counted directly on the agar) Or dissolved in a sterile solution such as peptone water or saline, so that aliquots can be aged in various media.
【０００４】 ドイツ連邦共和国特許公報第１１７３６４０号明細書は一般的に、製造時にゼ
ラチン膜の組織内に微生物を育成するための栄養物、さらに緩衝作用のある物質
、色素または微量の重金属または有害ガスのような空気中に含有された生物学的
に毒性作用のある物質を吸着することに適した化学薬品を浸潤することを提案し
ている。欠点はゼラチン膜に培養基または吸着物質を添加するにもかかわらず、
そこで収集された微生物から実際に汚染した培地中に当初存在していたものより
明らかに少ないコロニーしか増殖および同定できないことである。[0004] DE-A-1 173 640 generally discloses nutrients for the production of microorganisms in the tissue of the gelatin membrane during manufacture, as well as substances with a buffering effect, pigments or traces of heavy metals or harmful gases. It has been proposed to infiltrate chemicals suitable for adsorbing biologically toxic substances contained in the air, such as: The disadvantage is that despite the addition of culture media or adsorbents to the gelatin membrane,
Thus, the collected microorganisms can grow and identify significantly fewer colonies than were originally present in the contaminated medium.
【０００５】 従って本発明は、先行技術のゼラチン膜フィルタの場合よりも検出可能の微生
物数が明らかに増大している気体から微生物を収集するためのゼラチン膜フィル
タを作ることならびにこのゼラチン膜フィルタの製造方法を見出すことを目的と
している。Accordingly, the present invention is directed to making a gelatin membrane filter for collecting microorganisms from a gas having a significantly increased number of detectable microorganisms than is the case with prior art gelatin membrane filters, as well as the use of such gelatin membrane filters. The aim is to find a manufacturing method.
【０００６】 （発明の開示） この目的は独立請求項の構成要素を特徴とするゼラチン膜フィルタにより解決
される。この発明の好ましいさらなる態様は従属請求項の構成要素により挙げら
れている。[0006] This object is solved by a gelatin membrane filter featuring the features of the independent claims. Preferred further aspects of the invention are mentioned by the elements of the dependent claims.
【０００７】 驚くべきことに、ゼラチン膜フィルタがいわゆる浸透保護物質を含有する場合
、濾過により気体からゼラチン膜フィルタに収集された検出可能の微生物数が明
らかに増大することが見出された。この浸透保護物質には、たとえばイノシトー
ル、ベタイン、リシン、オキシノイリンおよび他のものが含まれる。これらの物
質は微生物の脱水、浸透ストレスに反対の作用をすることが知られている。明ら
かに微生物の一部はゼラチン膜フィルタに収集する際、細胞水においてこの微生
物がもはや生存能力がなくなり、熟成によりもはやコロニー形成を検出できない
ほどに失われる。この点については同数の細菌を与えられたゼラチン膜フィルタ
が、たとえば１分間の代りにさらに８分間７．５ｍ３／ｈの空気量により貫流さ
れた場合、残りは細菌の約半分だけが検出されることができたという事実も示唆
している。ゼラチン膜フィルタが微生物を明らかに湿らせて増殖能力を保持され
るべきであるため、この効果はゼラチン膜フィルタでは期待されたことがなかっ
た（案内書“実験室濾過、微生物学、電気泳動法”１４頁、ザルトリウス ゲー エムベーハー１９８４）。Surprisingly, it has been found that when the gelatin membrane filter contains a so-called osmoprotectant, the number of detectable microorganisms collected on the gelatin membrane filter from the gas by filtration is significantly increased. The osmoprotectants include, for example, inositol, betaine, lysine, oxyneurin and others. These substances are known to have the opposite effects on microbial dehydration and osmotic stress. Apparently, when some of the microorganisms are collected on a gelatin membrane filter, they are no longer viable in the cell water and are lost to maturity so that colony formation can no longer be detected. In this regard, if a gelatin membrane filter fed with the same number of bacteria is flowed through, for example, for 7.5 minutes instead of 1 minute with an air volume of 7.5 m 3 / h, only about half of the bacteria will be detected. It also suggests the fact that it was possible. This effect has never been expected with gelatin membrane filters, since gelatin membrane filters should clearly moisten microorganisms and retain their growth ability (see “Laboratory Filtration, Microbiology, Electrophoresis”). "Page 14, Sartorius G. M. Behr 1984).
【０００８】 この発明の好ましい一実施態様において、ゼラチン膜フィルタが微生物を収集
するために１分間およびさらに８分間７．５ｍ３／ｈの空気流により貫流される
場合、浸透保護物質のないゼラチン膜フィルタに比べ生存可能の微生物が少なく
とも２倍の数になる量でゼラチン膜フィルタが浸透保護物質を含有する。つまり
、ゼラチン膜フィルタ内で同じ割合の浸透保護物質が、収集された同数の微生物
でゼラチン膜フィルタを貫流する気体の様々な量に対し生存可能の微生物の様々
な割合をもたらすことが見出された。In one preferred embodiment of the invention, the gelatin membrane filter without the osmoprotectant, when the gelatin membrane filter is flowed through with a 7.5 m 3 / h air flow for 1 minute and for a further 8 minutes to collect the microorganisms The gelatin membrane filter contains the osmoprotectant in an amount that results in at least twice as many viable microorganisms as the filter. That is, it has been found that the same proportion of osmoprotectant in a gelatin membrane filter results in different proportions of viable microorganisms for different amounts of gas flowing through the gelatin membrane filter with the same number of microorganisms collected. Was.
【０００９】 ゼラチン膜フィルタ内の浸透保護物質としての酢酸トリメチルアンモニウムの
濃度が、特に膜が得られた膜吸引溶液中にゼラチン濃度に対し０．００５から０
．７５％までの割合で存在している場合、特に適していることが証明された。膜
吸引溶液は、全膜吸引溶液に対し４．６から５．６％までの割合のゼラチンと、
全膜吸引溶液に対し３８から４６％までの割合のエタノールとを含有する水性の
均一な溶液である。それに加えて膜の安定性のために、ゼラチンが付加的に結合
剤により安定化されている場合に長所となる。この結合剤は、たとえばポリビニ
ルアルコールまたはデンプンとすることができる。[0009] The concentration of trimethylammonium acetate as a permeation protective substance in the gelatin membrane filter is preferably 0.005 to 0 relative to the gelatin concentration in the membrane suction solution from which the membrane was obtained.
. It has proven to be particularly suitable when present in proportions up to 75%. The membrane aspirating solution is composed of 4.6 to 5.6% of gelatin relative to the total membrane aspirating solution,
It is an aqueous homogeneous solution containing from 38 to 46% ethanol with respect to the total membrane suction solution. In addition, due to the stability of the membrane, it is advantageous if the gelatin is additionally stabilized by a binder. This binder can be, for example, polyvinyl alcohol or starch.
【００１０】 ゼラチンの構造に基づき浸透保護物質がゼラチンマトリックスを可能な限り均
一に浸潤する場合に長所となる。これらの物質がゼラチン膜マトリックスの外部
表面上にのみ存在する場合、長時間保管するとき浸透保護物質の一部が内部のマ
トリックスへ移動して、全微生物に対して制限された作用のみがあり、またはも
はや全く作用しなくなる危険がある。従って浸透保護物質は本発明による好まし
い方法においてゼラチン膜フィルタが作られる相反転過程を通して膜マトリック
スの中に均一な分布で組み込まれる。そのために浸透保護物質が膜吸引溶液の中
へ取り込まれる。選択肢として浸透保護物質がゼラチン膜フィルタの形成中に凝
固槽に取り込まれる。これは全溶液に対し１０から２０％までの割合のアルコー
ル、好ましくはメタノールを含有する酢酸メチルから成り、ならびに全溶液に対
し約２％までの濃度の浸透保護物質から成る第１の凝固槽内で行われる。この第
１の凝固槽内ではゼラチン膜フィルタがまだ完全に形成されていないため、その
結果、浸透保護物質を全ての膜マトリックスの中に浸潤することができる。It is advantageous if the osmoprotectant infiltrates the gelatin matrix as uniformly as possible, based on the structure of the gelatin. If these substances are only present on the outer surface of the gelatin membrane matrix, part of the osmotic protective substance migrates to the inner matrix when stored for a long time and has only a limited effect on all microorganisms, Or there is a danger that it will no longer work at all. Thus, the osmoprotectant is incorporated in a uniform distribution within the membrane matrix through the phase inversion process in which the gelatin membrane filter is made in the preferred method according to the invention. To that end, a osmoprotectant is incorporated into the membrane suction solution. Optionally, a osmoprotectant is incorporated into the coagulation bath during formation of the gelatin membrane filter. This consists in a first coagulation bath consisting of methyl acetate containing 10 to 20% of alcohol, preferably methanol, based on the total solution, and of a osmoprotectant at a concentration of about 2% based on the total solution. Done in In this first coagulation bath, the gelatin membrane filter has not yet been completely formed, so that the osmoprotectant can infiltrate into all the membrane matrix.
【００１１】 この発明によるゼラチン膜フィルタは、ドイツ連邦共和国特許公報第１１７３
６４０号明細書により得られるものも含めた先行技術のゼラチン膜フィルタに比
べ、その取扱いが明らかに改善される増大した機械的安定性を有するだけでなく
、このゼラチン膜フィルタが少なくとも係数２だけ大きくなる明らかにより高い
空気流量を提供する。そのため、このゼラチン膜フィルタは細菌収集のための高
い空気流量に特に適している。それにより、この発明によるゼラチン膜フィルタ
によって微生物を収集するための時間を低減し、またはゼラチン膜フィルタがそ
の作用において損われずまたは分解されることなく、より大きい容積を濾過する
ことができるため、非常にわずかに負荷された気体を試験することも可能である
。[0011] A gelatin membrane filter according to the present invention is disclosed in German Patent Publication No. 1173.
In addition to having increased mechanical stability whose handling is clearly improved compared to prior art gelatin membrane filters, including those obtained according to US Pat. No. 640, this gelatin membrane filter is at least two times larger. Provides a significantly higher air flow. Therefore, this gelatin membrane filter is particularly suitable for high air flow for bacterial collection. Thereby, it is possible to reduce the time for collecting microorganisms by the gelatin membrane filter according to the present invention, or to filter a larger volume without the gelatin membrane filter being damaged or degraded in its operation, It is also possible to test very slightly loaded gases.
【００１２】 この発明によるゼラチン膜フィルタの製造方法は、 ａ）少なくともゼラチンとエタノールとを含有する水性の均一な膜吸引溶液が製
造され、 ｂ）薄膜が膜吸引溶液から基床上に伸展され、 ｃ）この薄膜がゲル化相を形成するために空気にさらされ、 ｄ）このゲル化相が沈殿槽内で再処理するために取り込まれ、 ｅ）ゼラチン膜フィルタが乾燥される、 ことにより実施され、かつ ｆ）膜吸引溶液が浸透保護物質を少なくとも1つ添加されることを特徴とする。The method for producing a gelatin membrane filter according to the present invention comprises the steps of: a) producing an aqueous uniform membrane suction solution containing at least gelatin and ethanol; b) extending a thin film from the membrane suction solution onto a base bed; A) exposing the film to air to form a gelled phase; d) entraining the gelled phase for reprocessing in a sedimentation vessel; and e) drying the gelatin membrane filter. And f) the membrane suction solution is added with at least one osmoprotectant.
【００１３】 この方法の好ましい一実施態様において、膜吸引溶液はゼラチンに対し０．０
０５および０．７５％の間の酢酸トリメチルアンモニウムを含有する。In one preferred embodiment of the method, the membrane aspirating solution is 0.0
Contains between 05 and 0.75% trimethylammonium acetate.
【００１４】 この方法のもう１つの実施態様において、ゼラチン膜フィルタの機械的な安定
性を増大するために結合剤が全膜吸引溶液に対し０．０２から０．１％までの割
合で膜吸引溶液全体に取り込まれる。[0014] In another embodiment of the method, the binder is used to increase the mechanical stability of the gelatin membrane filter at a rate of 0.02 to 0.1% of the total membrane suction solution. Incorporated throughout the solution.
【００１５】 この発明のもう１つ別の好ましい実施態様において、全体の第１の凝固槽に対
し１０から２０％までの割合のアルコール、特にメタノールを含有する酢酸メチ
ルが第１の凝固槽として使用される。膜はこれが純酢酸メチルから成る第２の凝
固槽の中に移行される前に、室温で１時間から３時間までの持続時間にわたりこ
の第１の凝固槽内にとどまる。好ましくはガンマ線により乾燥および滅菌が接続
される。In another preferred embodiment of the invention, methyl acetate containing from 10 to 20% of alcohol, in particular methanol, is used as the first coagulation bath relative to the total first coagulation bath. Is done. The membrane stays in this first coagulation bath at room temperature for a duration of 1 to 3 hours before it is transferred into a second coagulation bath consisting of pure methyl acetate. Drying and sterilization are preferably connected by gamma radiation.
【００１６】 ここで、この発明は以下の実施例を利用して説明する。Here, the present invention will be described using the following embodiments.
【００１７】 例１ ゼラチン膜フィルタを製造するために、ゼラチン２００ｇ（ゼリタ粉末ゼラチ
ン２５０ブルーム、デーゲーエフ ドイチェ ゼラチン-ファブリケン シュトェー
ス アーゲー）とポリビニルアルコール２ｇ（Ｍｏｗｉｏｌタイプ１８−８８、 ヘキスト アーゲー）とが６０℃で１時間攪拌しながら水２０００ｇに溶解され 、それに続きエタノール１６４５ｇと酢酸トリメチルアンモニウム０．０２ｇと
が溶解された水１０ｇに混合される。３３℃に温度調節された膜吸引溶液が強さ
３５０μｍの薄膜に基床上で伸張されて、室温で５分間相対湿度４５％の空気に
さらされる。ゲル化された薄膜は基床と共にメタノール１４％の割合の酢酸メチ
ルから成る第１の凝固槽内で持続時間３時間のために、その後、純酢酸メチルか
ら成る第２の凝固槽内で持続時間３時間のために取り込まれる。ゼラチン膜フィ
ルタが基床から引抜かれて乾燥される。 このようにして得られたゼラチン膜フィルタは約３μｍの孔径と１３８ｌ／ｍ
ｉｎ ｃｍ２ｂａｒの空気流量とを有する。 例１に従って得られたゼラチン膜フィルタが試料Ａと呼ばれる。Example 1 To prepare a gelatin membrane filter, 200 g of gelatin (250 g of Zelita powdered gelatin, Degeev Deutsche Gelatin-Fabricen Stöger AG) and 2 g of polyvinyl alcohol (Mowiol type 18-88, Hoechst AG) at 60 ° C. The mixture is dissolved in 2000 g of water while stirring for 1 hour, and then mixed with 10 g of water in which 1645 g of ethanol and 0.02 g of trimethylammonium acetate are dissolved. A membrane suction solution, thermostated at 33 ° C., is stretched on a substrate into a 350 μm strength membrane and exposed to air at 45% relative humidity for 5 minutes at room temperature. The gelled film together with the base is durable for 3 hours in a first coagulation bath consisting of 14% methanol in methyl acetate and then in a second coagulation bath consisting of pure methyl acetate. Ingested for 3 hours. The gelatin membrane filter is pulled out of the base and dried. The gelatin membrane filter thus obtained has a pore size of about 3 μm and 138 l / m 2.
with an air flow rate of in cm 2 bar. The gelatin membrane filter obtained according to Example 1 is called Sample A.
【００１８】 例２−５ 例１に類似して試料Ｂ、Ｃ、ＤおよびＥが製造され、単に膜吸引溶液内の酢酸
トリメチルアンモニウムの質量のみが変化される（表、第２行目）。Examples 2-5 Samples B, C, D and E are prepared analogously to Example 1 and only the mass of trimethylammonium acetate in the membrane suction solution is changed (table, line 2).
【００１９】 例６ 例１に類似して比較試料Ｆが製造されるが、ただし酢酸トリメチルアンモニウ
ムの添加が省略される。Example 6 A comparative sample F is prepared analogously to Example 1, except that the addition of trimethylammonium acetate is omitted.
【００２０】 浸透保護物質の効果を検査するために、例１ないし６に従って製造されたゼラ
チン膜フィルタＡないしＦが１分間同量のエッシャリキア コリＡＴＣＣ８７３ ９の細菌を与えられて、さらに８分間にわたり流量７．５ｍ３／ｈの空気が与え
られたゼラチン膜フィルタを通して吸引された。ゼラチン膜フィルタはその直後
にトリプトン大豆肉汁寒天（ＴＳＢＡ培養肉汁）を含有する寒天培養厚紙プレー
トに塗布されて、夜通し３７℃に保温培養されて、コロニー数が顕微鏡で同定さ
れた。To test the effect of the osmoprotectant, gelatin membrane filters A to F prepared according to Examples 1 to 6 were fed with the same amount of bacteria of Escherichia coli ATCC 873 9 for 1 minute and flowed for an additional 8 minutes. 7.5 m 3 / h of air were aspirated through the provided gelatin membrane filter. Immediately thereafter, the gelatin membrane filter was applied to an agar culture cardboard plate containing trypton soybean broth agar (TSBA culture broth), incubated at 37 ° C. overnight, and the number of colonies was identified with a microscope.
【００２１】 この結果は表の第３列および第４列に掲載した。この結果はそれぞれ５回の測
定からの平均値を表しており、浸透保護物質としての酢酸トリメチルアンモニウ
ムが、膜吸引溶液中のゼラチン濃度に対し０．０１から０．２５％までの割合で
存在していて、ゼラチン膜フィルタが微生物を収集するために１分間およびさら
に８分間７．５ｍ３／ｈの空気流により貫流される場合、浸透保護物質のないゼ
ラチン膜フィルタ（試料Ｆ）に比べ生存可能の微生物が少なくとも２倍の数にな
る（試料Ａ、ＢおよびＣ）ことを示している。The results are listed in the third and fourth columns of the table. The results each represent the average value from five measurements, in which trimethylammonium acetate as the osmotic protective substance was present at a ratio of 0.01 to 0.25% based on the gelatin concentration in the membrane suction solution. And is more viable than a gelatin membrane filter without osmotic protective material (Sample F) when the gelatin membrane filter is flowed through with a 7.5 m 3 / h air flow for 1 minute and an additional 8 minutes to collect microorganisms Of microorganisms at least doubled (samples A, B and C).
【００２２】 表Table
【表１】 [Table 1]
【００２３】 試験材料および試験方法 試験細菌： この試験にはエッシャリキア コリＡＴＴＣＣ８７３９が使用された。この菌 株は製剤の有効性試験のための微生物学品質保証課で広く使用されている。１ｍ
ｌあたり約１０３コロニー形成単位［ＫＢＥ／ｍｌ］の濃度を有する凍結された
フラスコが使用された。それぞれ解凍された懸濁液０．１ｍｌが同一の２つの対
比でトリプトン大豆肉汁寒天（ＴＳＢＡ培養肉汁）を含むプレートに塗抹された
。寒天プレートが夜通し３７℃±２℃で熟成され、細菌がグラム染色法により顕
微鏡で同定された。この対象担体が保存見本として資料作成された。Test Materials and Test Methods Test Bacteria: Escherichia coli ATTCC 8739 was used for this test. This strain is widely used in the Microbiology Quality Assurance Division for testing the efficacy of pharmaceuticals. 1m
Frozen flask having a concentration of about 103 colony forming units per l [KBE / ml] was used. 0.1 ml of each thawed suspension was smeared onto a plate containing tryptone soy broth agar (TSBA culture broth) in two identical contrasts. Agar plates were aged at 37 ° C. ± 2 ° C. overnight and bacteria were identified microscopically by Gram staining. This target carrier was prepared as a storage sample.
【００２４】 培養懸濁液が受容能２５０ｍｌを有する容器内にあらかじめプレート上で洗浄
されたコロニーを含有する上記培養肉汁５０ｍｌの接種により製造された。この
容器が夜通し３７℃±２℃で振盪しながら水槽内で熟成された。この懸濁液が培
養技術を利用して検査され、滅菌フラスコの中に移し替えられて、別の使用まで
４℃±２℃で保管された。A culture suspension was prepared by inoculating 50 ml of the above mentioned broth containing colonies, which had been previously washed on a plate, into a container having a capacity of 250 ml. The vessel was aged in a water bath with shaking at 37 ° C. ± 2 ° C. overnight. This suspension was examined using culture techniques, transferred into sterile flasks, and stored at 4 ° C. ± 2 ° C. until further use.
【００２５】 バチルス スブチリス ヴァル ニガー（ＮＣＴＣ１００７３）の芽胞が微生物 学上のトレーサとして使用された。この芽胞が遠心分離器によって３回洗浄され
、滅菌蒸留水で再び懸濁液にされた。バチルス スブチリスの菌株溶液の濃度は 約２ｘ１０１１ＫＢＥ／ｍｌであった。バチルス スブチリスから成る１リット ル懸濁液は、菌株溶液が約２５ｘ１０９ＫＢＥ／ｍｌに滅菌蒸留水で希釈され、
４０分間６０℃で場合により存在する植物性の有機体を壊死させるために加熱さ
れた。この試験に使用された芽胞懸濁液の同定はバチルス スブチリス芽胞２． ２６ｘ１０９ＫＢＥ／ｍｌの最終濃度が生じた。Spores of Bacillus subtilis val niger (NCTC 10073) were used as microbiological tracers. The spores were washed three times by a centrifuge and resuspended with sterile distilled water. The concentration of the Bacillus subtilis strain solution was about 2 × 10 11 KBE / ml. A one liter suspension of Bacillus subtilis was prepared by diluting the bacterial solution to approximately 25 × 10 9 KBE / ml with sterile distilled water.
Heated for 40 minutes at 60 ° C. to necrotize any plant organisms present. The spore suspension used in this study was identified as Bacillus subtilis spores. A final concentration of 26 × 10 9 KBE / ml resulted.
【００２６】 霧化懸濁液の準備： エッシャリキア コリＡＴＣＣ８７３９の懸濁液が使用前に２週間４℃±２℃ で保管された。培養技術を利用した再同定は、エッシャリキア コリ濃度がこの 期間中一定にとどまっていることを示した。導入される試験は最終試験に適した
濃度を突止めるため種々の試験およびトレーサ細菌の濃度で実施された。Preparation of Atomized Suspension: A suspension of Escherichia coli ATCC 8739 was stored at 4 ° C. ± 2 ° C. for 2 weeks before use. Re-identification using culture techniques showed that the concentration of Escherichia coli remained constant during this period. The tests introduced were performed at various tests and concentrations of tracer bacteria to determine the appropriate concentration for the final test.
【００２７】 エッシャリキア コリＡＴＣＣ８７３９の菌株懸濁液の濃度は２．１ｘ１０１ ０ ＫＢＥ／ｍｌであった。霧化懸濁液１０ｍｌの製造はエッシャリキア コリ５ ｍｌ懸濁液（蒸留水で希釈して、７．６ｘ１０３ないし２．８ｘ１０４ＫＢＥ／
ｍｌを得る）とバチルス スブチリス５ｍｌ懸濁液（蒸留水で希釈して、４．０ ｘ１０３ＫＢＥ／ｍｌを得る）との混合により行われた。最終懸濁液が３０分以
内に霧化された（エーロゾル化）。The concentration of the Escherichia coli ATCC 8739 suspension was 2.1 × 10 10 KBE / ml. The production of 10 ml of the nebulized suspension is made up of a 5 ml suspension of Escherichia coli (diluted with distilled water to give 7.6 × 10 3 to 2.8 × 10 4 KBE /
ml) and a 5 ml suspension of Bacillus subtilis (diluted with distilled water to obtain 4.0 × 10 3 KBE / ml). The final suspension atomized within 30 minutes (aerosolization).
【００２８】 エーロゾル試験台： 試験台は閉鎖および密閉されたチャンバ内で細菌エーロゾルを発生するための
コリソン型３ジェットネブライザの霧化器から成る。エーロゾルチャンバは内径
４５ｍｍを有する特殊鋼製の長さ１ｍの管系と連結されている。ザルトリウス社
製の空気収集器のフィルタヘッドはこの配管に対応するアダプタを利用して接続
されている。チャンバの入口は高性能の空気フィルタにより保護されている。Aerosol test bench: The test bench consists of a Collison-type three-jet nebulizer atomizer for generating bacterial aerosol in a closed and sealed chamber. The aerosol chamber is connected to a 1 m long tubing made of special steel having an inner diameter of 45 mm. The filter head of the Sartorius air collector is connected using an adapter corresponding to this pipe. The inlet of the chamber is protected by a sophisticated air filter.
【００２９】 検定証明書、チューブおよびフィルタヘッド付きのザルトリウスＭＤ８エアス
キャン空気細菌収集器（シリーズ番号第９６０７００１号）は取扱説明書に従っ
て操作された。A Sartorius MD8 airscan air bacteria collector with certificate of certification, tubing and filter head (Series No. 9607001) was operated according to the instruction manual.
【００３０】 ゼラチン膜フィルタ： それぞれ試料Ａ、Ｂ、Ｃ、Ｄ、ＥおよびＦとして表示された小孔サイズ約３．
０μｍを有する滅菌ゼラチン膜フィルタが使用された。改質されたゼラチン膜フ
ィルタＡ、Ｂ、Ｃ、ＤおよびＥは上記の表に挙げた様々な量の酢酸トリメチルア
ンモニウムを浸透保護物質として含有した。Gelatin Membrane Filter: Approximately 3. pore size indicated as Samples A, B, C, D, E and F, respectively.
A sterile gelatin membrane filter with 0 μm was used. Modified gelatin membrane filters A, B, C, D and E contained various amounts of trimethylammonium acetate as listed in the table above as a osmoprotectant.
【００３１】 試験方法： 霧化されるべき懸濁液９ｍｌがコリソン型霧化器の中に装入された（１ｍｌが
微生物学的検出のために残された）。各々のゼラチン膜フィルタがフィルタヘッ
ドの中に置かれ、試験台に（上述のように）ねじ止めされた。Test method: 9 ml of the suspension to be atomized were charged in a Collison atomizer (1 ml was left for microbiological detection). Each gelatin membrane filter was placed in the filter head and screwed to the test bench (as described above).
【００３２】 ＭＤ８エアスキャンが空気流量７．５ｍ３／ｈに設定された。ＭＤ８エアスキ
ャンがスイッチ投入されて、１５秒後にコリソン型霧化器が圧縮ガス管１８０ｋ
Ｐａに接続されることにより活性化された。この霧化器が１分間活性化されて、
その後ＭＤ８がオフにされた。The MD8 air scan was set at an air flow of 7.5 m 3 / h. The MD8 air scan is switched on, and 15 seconds later, the Collison atomizer turns on the compressed gas pipe 180k.
It was activated by being connected to Pa. This atomizer is activated for 1 minute,
Then MD8 was turned off.
【００３３】 ゼラチン膜フィルタが細菌を含有するエーロゾル（空気細菌収集器ＭＤ８を利
用して吸引）を１分間の付与後に直接的に、または濾過された空気をさらに８分
間付与後にＴＳＢＡプレートに載置され、さらに上述のように熟成および測定さ
れた。The gelatin membrane filter is placed on the TSBA plate directly after application of the aerosol containing bacteria (aspirated using the air bacteria collector MD8) for 1 minute or after application of the filtered air for a further 8 minutes. And further aged and measured as described above.
【００３４】 この発明の説明で使用した％表示は質量百分率である。The percentages used in the description of the invention are percentages by mass.
【手続補正書】特許協力条約第３４条補正の翻訳文提出書[Procedural Amendment] Submission of translation of Article 34 Amendment of the Patent Cooperation Treaty
【提出日】平成１１年１０月１９日（１９９９．１０．１９）[Submission date] October 19, 1999 (1999.10.19)
【手続補正１】[Procedure amendment 1]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】特許請求の範囲[Correction target item name] Claims
【補正方法】変更[Correction method] Change
【補正内容】[Correction contents]
【特許請求の範囲】[Claims]
【手続補正２】[Procedure amendment 2]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】０００４[Correction target item name] 0004
【補正方法】変更[Correction method] Change
【補正内容】[Correction contents]
【０００４】 ＷＯ９２０８３５５Ａは細胞脱水に基づく微生物としての細菌の損傷を防止す
る浸透保護物質としてベタイン、トレハロース、プロリン、コリン、グルタミン
酸塩、グルタミンおよびγ-アミノ酪酸のような小さい有機分子について言及し ている。 ドイツ連邦共和国特許公報第１１７３６４０号明細書は一般的に、製造時にゼ
ラチン膜の組織内に微生物を育成するための栄養物、さらに緩衝作用のある物質
、色素または微量の重金属または有害ガスのような空気中に含有された生物学的
に毒性作用のある物質を吸着することに適した化学薬品を浸潤することを提案し
ている。欠点はゼラチン膜に培養基または吸着物質を添加するにもかかわらず、
そこで収集された微生物から実際に汚染した培地中に当初存在していたものより
明らかに少ないコロニーしか増殖および同定できないことである。[0004] WO 9208355 A mentions small organic molecules such as betaine, trehalose, proline, choline, glutamate, glutamine and γ-aminobutyric acid as osmoprotectants which prevent bacterial damage as a microorganism based on cell dehydration. . DE-A-1 173 640 generally describes nutrients for the production of microorganisms in the tissue of the gelatin membrane during manufacture, as well as substances having a buffering action, pigments or traces of heavy metals or noxious gases. It proposes infiltrating chemicals suitable for adsorbing biologically toxic substances contained in the air. The disadvantage is that despite the addition of culture media or adsorbents to the gelatin membrane,
Thus, the collected microorganisms can grow and identify significantly fewer colonies than were originally present in the contaminated medium.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 ヤスヒホフ， ヘルムート ドイツ連邦共和国 Ｄ−17489 グライフ スヴァルト、ゲルハルト−カッチュ−シュ トラーセ 14 (72)発明者 トゥ ヴィン， クゥオン ドイツ連邦共和国 Ｄ−31167 ボッケネ ム、プロフ．−マルティーニ−シュトラー セ 16 Ｆターム(参考） 4B029 AA09 HA10 4D006 GA44 KE01R KE28R MA03 MC33X MC69X MC90 NA05 NA16 NA17 NA40 PB17 PB24 PC41 ────────────────────────────────────────────────── ─── Continuing on the front page (72) Inventor Jaschoff, Helmut Germany D-17489 Greif Sward, Gerhard-Kacchustrasse 14 (72) Inventor Tu Vinh, Kuon Germany D-31167 Bokkenem, Prof . -Martini-Strasse 16 F term (reference) 4B029 AA09 HA10 4D006 GA44 KE01R KE28R MA03 MC33X MC69X MC90 NA05 NA16 NA17 NA40 PB17 PB24 PC41
Claims (13)
て、 ゼラチン膜フィルタが浸透保護物質を含有することを特徴とする、前記ゼラチン
膜フィルタ。1. A gelatin membrane filter for collecting microorganisms from a gas, wherein the gelatin membrane filter contains a permeable protective substance.
ないゼラチン膜フィルタに比べ生存可能の微生物が少なくとも２倍の数になる量
でゼラチン膜フィルタが浸透保護物質を含有することを特徴とする、請求項１に
記載のゼラチン膜フィルタ。2. When the gelatin membrane filter is flowed through with a 7.5 m 3 / h air flow for 1 minute and also 8 minutes to collect microorganisms, it is more viable than a gelatin membrane filter without osmotic protective substance. The gelatin membrane filter according to claim 1, characterized in that the gelatin membrane filter contains an osmotic protective substance in an amount to at least double the number of microorganisms.
ンモニウムを含有することを特徴とする、請求項１又は２に記載のゼラチン膜フ
ィルタ。3. The gelatin membrane filter according to claim 1, wherein the gelatin membrane filter contains trimethylammonium acetate as a permeation protective substance.
請求項１〜３にのいずれかに記載のゼラチン膜フィルタ。4. The gelatin membrane filter contains a binder,
The gelatin membrane filter according to claim 1.
７．５ｍ３／ｈの空気流により貫流される場合、この浸透保護物質のないゼラチ
ン膜フィルタに比べ生存可能の微生物が少なくとも２倍の数になる全膜吸引溶液
に対する割合の少なくとも1つの浸透保護物質、 ｄ）全膜吸引溶液に対し０．０２から０．１％までの割合の結合剤、 から成る水性の均一な膜吸引性溶液から得られる請求項１〜４のいずれかに記載
のゼラチン膜フィルタ。5. The following components by phase inversion: a) gelatin in a proportion of 4.6 to 5.6% of the total membrane suction solution, b) proportion of 38 to 46% of the total membrane suction solution. Ethanol, c) Viable microorganisms compared to gelatin membrane filters without this osmoprotectant, when the gelatin membrane filter is flowed through with a 7.5 m 3 / h air flow for 1 minute and an additional 8 minutes to collect the microorganisms At least one osmoprotectant in a ratio to the total membrane aspirating solution, which is at least twice as high; d) a binder in a proportion of 0.02 to 0.1% of the total membrane aspirating solution; The gelatin membrane filter according to any one of claims 1 to 4, which is obtained from a suitable membrane suction solution.
．００５から０．７５％までの割合の酢酸トリメチルアンモニウムが存在するこ
とを特徴とする、請求項５に記載のゼラチン膜フィルタ。6. The osmotic protective substance in the membrane aspirating solution has a concentration of 0% relative to the concentration of gelatin.
. A gelatin membrane filter according to claim 5, characterized in that a proportion of trimethylammonium acetate of from 005 to 0.75% is present.
方法であって、 ａ）少なくともゼラチンとエタノールとを含有する水性の均一な膜吸引溶液が
製造され、 ｂ）薄膜が膜吸引溶液から基床上に伸展され、 ｃ）この薄膜がゲル化相を形成するために空気にさらされ、 ｄ）このゲル化相が沈殿槽内で再処理するために取り込まれ、 ｅ）ゼラチン膜フィルタが乾燥される、 ことにより、 ｆ）膜吸引溶液に浸透保護物質が少なくとも1つ添加されることを特徴とする 、前記方法。7. The method for producing a gelatin membrane filter according to claim 1, wherein: a) an aqueous uniform membrane suction solution containing at least gelatin and ethanol is produced; b) a thin film Is spread on the substrate from the membrane suction solution, c) the film is exposed to air to form a gelled phase, d) the gelled phase is entrained for reprocessing in a settling tank, e) Drying the gelatin membrane filter, f) adding at least one osmoprotectant to the membrane suction solution.
ｈの空気流により１分間およびさらに８分間貫流される場合、この浸透保護物質
のないゼラチン膜フィルタに比べ生存可能の微生物が少なくとも２倍の数になる
量で膜吸引溶液の浸透保護物質が添加されることを特徴とする、請求項７に記載
のゼラチン膜フィルタの製造方法。8. A gelatin membrane filter for collecting microorganisms at 7.5 m 3 /
h and 1 minute and for a further 8 minutes, the osmoprotectant of the membrane aspirating solution is added in an amount which at least doubles the number of viable microorganisms compared to this membrane filter without osmoprotectant. The method for producing a gelatin membrane filter according to claim 7, wherein the method is performed.
液に加えるゼラチンの濃度に対し０．００５から０．７５％までの割合で添加さ
れることを特徴とする、請求項７または８に記載のゼラチン膜フィルタの製造方
法。9. The method according to claim 7, wherein trimethylammonium acetate is added as a osmotic protective substance at a ratio of 0.005 to 0.75% with respect to the concentration of gelatin added to the membrane suction solution. A method for producing the gelatin membrane filter according to the above.
割合で添加されることを特徴とする、請求項７〜９のいずれかに記載のよるゼラ
チン膜フィルタの製造方法。10. The gelatin membrane filter according to claim 7, wherein the binder is added in a ratio of 0.02 to 0.1% with respect to the whole membrane suction solution. Manufacturing method.
好ましくはメタノール１０から２０％までの割合を含有する酢酸メチルから成る
第１の凝固槽の中に取り込まれることを特徴とする、請求項７〜１０のいずれか
に記載の方法。11. The gelling phase according to step d) is treated with alcohol for reprocessing,
11. Process according to claim 7, characterized in that it is taken up in a first coagulation bath, preferably consisting of methyl acetate containing a proportion of from 10 to 20% of methanol.
均一な膜吸引溶液が製造され、 ｂ）薄膜が膜吸引溶液から基床上に伸展され、 ｃ）この薄膜がゲル化相を形成するために空気にさらされ、 ｄ）このゲル化相が沈殿槽内で再処理するために取り込まれ、 ｅ）ゼラチン膜フィルタが乾燥される、 ことにより、 工程ｄ）に従ってゲル化相が酢酸メチルと、アルコールと、好ましくは１０か
ら２０％までの割合のメタノールと、浸透保護物質として第１の凝固槽の全溶液
に対し２％未満の割合の酢酸トリメチルアンモニウムとから成る第１の凝固槽の
中で再処理されることを特徴とする、請求項１〜６のいずれかに記載のゼラチン
膜フィルタの製造方法。12. A) An aqueous uniform membrane suction solution containing at least gelatin and ethanol is prepared; b) A thin film is spread on the substrate from the membrane suction solution; c) The thin film forms a gelled phase D) the gelled phase is taken up for reprocessing in a settling tank, e) the gelatin membrane filter is dried, whereby the gelled phase is methyl acetate according to step d) Of a first coagulation bath comprising alcohol, preferably methanol in a proportion of from 10 to 20%, and trimethylammonium acetate in a proportion of less than 2%, based on the total solution in the first coagulation tank, as osmotic protective substance. The method for producing a gelatin membrane filter according to any one of claims 1 to 6, wherein the gelatin membrane filter is reprocessed in water.
添加されることを特徴とする、請求項１２に記載の方法。13. The method according to claim 12, wherein the binder is added to the membrane suction solution in a proportion of 0.02 to 0.1%.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19750215.6 | 1997-11-13 | ||
| DE19750215A DE19750215C1 (en) | 1997-11-13 | 1997-11-13 | Gelatin membrane filter increasing number of viable micro-organisms collected from gas |
| PCT/EP1998/007118 WO1999025465A1 (en) | 1997-11-13 | 1998-11-07 | Gelatine membrane filters and method for producing the same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2001523549A true JP2001523549A (en) | 2001-11-27 |
Family
ID=7848554
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2000520893A Pending JP2001523549A (en) | 1997-11-13 | 1998-11-07 | Gelatin membrane filter and method for producing the same |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP1019179A1 (en) |
| JP (1) | JP2001523549A (en) |
| DE (1) | DE19750215C1 (en) |
| WO (1) | WO1999025465A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009139115A (en) * | 2007-12-04 | 2009-06-25 | Hitachi Plant Technologies Ltd | Capturing device and analysis system using the same |
| JP2010528643A (en) * | 2007-06-07 | 2010-08-26 | ビオメリュー | A device that dissolves microorganisms present in the environment or clinical samples and extracts nucleic acids from the microorganisms for analysis |
| JP5263290B2 (en) * | 2008-06-27 | 2013-08-14 | 株式会社日立プラントテクノロジー | Bacteria collection carrier cartridge, carrier treatment apparatus, and bacteria measurement method |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19814715A1 (en) * | 1998-04-02 | 1999-10-07 | Sartorius Gmbh | Gelatin membrane filters, process for their preparation and their use |
| DE60220989T2 (en) * | 2001-02-05 | 2008-03-13 | Millipore Corp., Billerica | DETECTION OF MICROORGANISMS |
| GB2387130A (en) | 2002-04-04 | 2003-10-08 | Fluid Technologies Plc | Hollow fibre filter membrane unit with microorganism detector, and associated usage |
| DE102014203855A1 (en) * | 2014-03-03 | 2015-09-03 | Biotec Gmbh, Umwelt-Analytik-Beratung-Service | Application for PCR analysis of airborne nucleic acids |
| EP3336515B1 (en) | 2016-12-14 | 2022-05-11 | Airbus Defence and Space GmbH | Hand-held air sampler, including filter magazine for storing and positioning filter elements |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE1173640B (en) * | 1959-05-21 | 1964-07-09 | Membranfiltergesellschaft G M | Process for the production of membrane filters |
| US5695541A (en) * | 1990-11-13 | 1997-12-09 | Liphatech, Inc. | Process for preparation of bacterial agricultural products |
| US5780709A (en) * | 1993-08-25 | 1998-07-14 | Dekalb Genetics Corporation | Transgenic maize with increased mannitol content |
| CA2221411A1 (en) * | 1995-05-19 | 1996-11-21 | Phytera, Inc. | Manipulation of plant cell and tissue cultures |
-
1997
- 1997-11-13 DE DE19750215A patent/DE19750215C1/en not_active Expired - Fee Related
-
1998
- 1998-11-07 WO PCT/EP1998/007118 patent/WO1999025465A1/en not_active Application Discontinuation
- 1998-11-07 JP JP2000520893A patent/JP2001523549A/en active Pending
- 1998-11-07 EP EP98955551A patent/EP1019179A1/en not_active Withdrawn
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010528643A (en) * | 2007-06-07 | 2010-08-26 | ビオメリュー | A device that dissolves microorganisms present in the environment or clinical samples and extracts nucleic acids from the microorganisms for analysis |
| JP2009139115A (en) * | 2007-12-04 | 2009-06-25 | Hitachi Plant Technologies Ltd | Capturing device and analysis system using the same |
| JP5263290B2 (en) * | 2008-06-27 | 2013-08-14 | 株式会社日立プラントテクノロジー | Bacteria collection carrier cartridge, carrier treatment apparatus, and bacteria measurement method |
| US9834806B2 (en) | 2008-06-27 | 2017-12-05 | Hitachi Plant Services Co., Ltd. | Microbe-collecting carrier cartridge, carrier treating apparatus, and method of measuring microbes |
Also Published As
| Publication number | Publication date |
|---|---|
| EP1019179A1 (en) | 2000-07-19 |
| WO1999025465A1 (en) | 1999-05-27 |
| DE19750215C1 (en) | 1999-02-04 |
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