CN110846375B - Culture medium composition for bacterial resistance counting, preparation method and application thereof - Google Patents
Culture medium composition for bacterial resistance counting, preparation method and application thereof Download PDFInfo
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- CN110846375B CN110846375B CN201911012056.8A CN201911012056A CN110846375B CN 110846375 B CN110846375 B CN 110846375B CN 201911012056 A CN201911012056 A CN 201911012056A CN 110846375 B CN110846375 B CN 110846375B
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- culture medium
- medium composition
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- concentration
- bacteria
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- 239000001963 growth medium Substances 0.000 title claims abstract description 77
- 239000000203 mixture Substances 0.000 title claims description 102
- 238000002360 preparation method Methods 0.000 title claims description 19
- 206010034133 Pathogen resistance Diseases 0.000 title claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 31
- 210000004556 brain Anatomy 0.000 claims abstract description 22
- 238000001802 infusion Methods 0.000 claims abstract description 21
- 238000001914 filtration Methods 0.000 claims description 40
- 238000004140 cleaning Methods 0.000 claims description 18
- 238000005303 weighing Methods 0.000 claims description 11
- 239000011148 porous material Substances 0.000 claims description 8
- 239000000126 substance Substances 0.000 claims description 7
- 239000006166 lysate Substances 0.000 claims 2
- 241000894006 Bacteria Species 0.000 abstract description 62
- 230000001580 bacterial effect Effects 0.000 abstract description 47
- 238000000034 method Methods 0.000 abstract description 47
- 239000002609 medium Substances 0.000 abstract description 16
- 238000012258 culturing Methods 0.000 abstract description 13
- 244000005700 microbiome Species 0.000 abstract description 8
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- 238000005054 agglomeration Methods 0.000 abstract description 3
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- 230000000996 additive effect Effects 0.000 description 21
- 239000002904 solvent Substances 0.000 description 21
- 230000002779 inactivation Effects 0.000 description 18
- 239000013028 medium composition Substances 0.000 description 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- 238000012360 testing method Methods 0.000 description 13
- 239000000725 suspension Substances 0.000 description 11
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 10
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 9
- 235000019796 monopotassium phosphate Nutrition 0.000 description 9
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 9
- 238000004659 sterilization and disinfection Methods 0.000 description 9
- 238000012986 modification Methods 0.000 description 8
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- 239000011780 sodium chloride Substances 0.000 description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- 230000036512 infertility Effects 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 5
- 241000191967 Staphylococcus aureus Species 0.000 description 5
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- 241000193830 Bacillus <bacterium> Species 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
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- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 241001478240 Coccus Species 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000834 fixative Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 241000588722 Escherichia Species 0.000 description 2
- 241000589989 Helicobacter Species 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 241000193998 Streptococcus pneumoniae Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- TZCXTZWJZNENPQ-UHFFFAOYSA-L barium sulfate Chemical compound [Ba+2].[O-]S([O-])(=O)=O TZCXTZWJZNENPQ-UHFFFAOYSA-L 0.000 description 2
- -1 but not limited to Chemical class 0.000 description 2
- 229920002301 cellulose acetate Polymers 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 230000000415 inactivating effect Effects 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- TZIHFWKZFHZASV-UHFFFAOYSA-N methyl formate Chemical compound COC=O TZIHFWKZFHZASV-UHFFFAOYSA-N 0.000 description 2
- 239000013618 particulate matter Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 229940031000 streptococcus pneumoniae Drugs 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 238000004506 ultrasonic cleaning Methods 0.000 description 2
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 1
- DURPTKYDGMDSBL-UHFFFAOYSA-N 1-butoxybutane Chemical compound CCCCOCCCC DURPTKYDGMDSBL-UHFFFAOYSA-N 0.000 description 1
- XFJQZJBQLJFFRR-UHFFFAOYSA-N 2,3,4-trichloro-5,6-dihydrobenzo[b][1]benzoxepine Chemical compound ClC=1C(=C(C2=C(C1)OC1=C(C=CC=C1)CC2)Cl)Cl XFJQZJBQLJFFRR-UHFFFAOYSA-N 0.000 description 1
- 241000589291 Acinetobacter Species 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 241000304886 Bacilli Species 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 1
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 241000588697 Enterobacter cloacae Species 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 241000194032 Enterococcus faecalis Species 0.000 description 1
- 241000589565 Flavobacterium Species 0.000 description 1
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 1
- 241000588915 Klebsiella aerogenes Species 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- 241000158728 Meliaceae Species 0.000 description 1
- 241000192041 Micrococcus Species 0.000 description 1
- KWYHDKDOAIKMQN-UHFFFAOYSA-N N,N,N',N'-tetramethylethylenediamine Chemical compound CN(C)CCN(C)C KWYHDKDOAIKMQN-UHFFFAOYSA-N 0.000 description 1
- 241000588653 Neisseria Species 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 241000588912 Pantoea agglomerans Species 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 239000004695 Polyether sulfone Substances 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 241000588768 Providencia Species 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000607720 Serratia Species 0.000 description 1
- 241000607715 Serratia marcescens Species 0.000 description 1
- 241000607768 Shigella Species 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 241001147736 Staphylococcus capitis Species 0.000 description 1
- 241000191963 Staphylococcus epidermidis Species 0.000 description 1
- 241000191984 Staphylococcus haemolyticus Species 0.000 description 1
- 241001464905 Staphylococcus saccharolyticus Species 0.000 description 1
- 108700025259 Streptococcus beta Proteins 0.000 description 1
- 241000194049 Streptococcus equinus Species 0.000 description 1
- 241000193996 Streptococcus pyogenes Species 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- WDIHJSXYQDMJHN-UHFFFAOYSA-L barium chloride Chemical compound [Cl-].[Cl-].[Ba+2] WDIHJSXYQDMJHN-UHFFFAOYSA-L 0.000 description 1
- 229910001626 barium chloride Inorganic materials 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- QGJOPFRUJISHPQ-NJFSPNSNSA-N carbon disulfide-14c Chemical compound S=[14C]=S QGJOPFRUJISHPQ-NJFSPNSNSA-N 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- IEJIGPNLZYLLBP-UHFFFAOYSA-N dimethyl carbonate Chemical compound COC(=O)OC IEJIGPNLZYLLBP-UHFFFAOYSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940092559 enterobacter aerogenes Drugs 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 239000012526 feed medium Substances 0.000 description 1
- 239000011152 fibreglass Substances 0.000 description 1
- 239000008098 formaldehyde solution Substances 0.000 description 1
- 239000013022 formulation composition Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
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- 238000007654 immersion Methods 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229910000358 iron sulfate Inorganic materials 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 235000011147 magnesium chloride Nutrition 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- XTAZYLNFDRKIHJ-UHFFFAOYSA-N n,n-dioctyloctan-1-amine Chemical compound CCCCCCCCN(CCCCCCCC)CCCCCCCC XTAZYLNFDRKIHJ-UHFFFAOYSA-N 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920006393 polyether sulfone Polymers 0.000 description 1
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 1
- 239000004810 polytetrafluoroethylene Substances 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000001223 reverse osmosis Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 235000011008 sodium phosphates Nutrition 0.000 description 1
- 238000013190 sterility testing Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- IMFACGCPASFAPR-UHFFFAOYSA-N tributylamine Chemical compound CCCCN(CCCC)CCCC IMFACGCPASFAPR-UHFFFAOYSA-N 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
- 239000010981 turquoise Substances 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 239000012856 weighed raw material Substances 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 229960001763 zinc sulfate Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/045—Culture media therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The present invention provides a medium comprising: MH broth at a concentration of 0.5 g/l to 40g/l, brain heart infusion at a concentration of 0.5 g/l to 60 g/l, and water. The technical scheme of the invention has the characteristics of accelerating the growth of microorganisms, reducing the background and reducing the occurrence of bacterial agglomeration growth, and the culture medium is applied to the counting of bacteria by a resistance method, so that the function of rapidly culturing the bacteria is achieved on one hand, and the requirements of reducing the background, namely reducing particulate matters, required by the resistance counting are met on the other hand.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to a culture medium, especially a bacterial culture medium.
Background
The culture medium is an in-vitro enrichment culture reagent, provides proper nutrition components and growth environment for the growth and propagation of microorganisms and accelerates the growth speed of the microorganisms. At present, the culture medium in domestic market has tens and various types, and can meet the requirements of culturing common clinical microorganisms. However, if the medium is used as a medium for counting bacteria by a resistance method, on one hand, the function of rapidly culturing bacteria is achieved, the background reduction required by resistance counting, namely the requirement of reducing particulate matters is also met, the medium on the market at present only meets the requirement of culturing common bacteria, and the requirement of the medium for counting bacteria by the resistance method cannot be met, so the medium for meeting the requirement of resistance counting is required.
Disclosure of Invention
The embodiment of the invention provides a bacterial culture medium used in a bacterial counting process by a resistance method, which at least solves the technical problems of slower bacterial culture and higher cost in the prior art.
According to an embodiment of the present invention, there is provided a medium composition for bacterial resistance counting, the medium composition comprising: MH broth at a concentration of 0.5 g/l to 40g/l, brain heart infusion at a concentration of 0.5 g/l to 60 g/l, and water.
Optionally, the above-described culture medium composition comprises: MH broth at a concentration of 5g/l to 27g/l, brain heart infusion at a concentration of 5g/l to 40g/l, and water.
Alternatively, the MH broth has a concentration of 21g/L, the brain heart infusion has a concentration of 12g/L, and the water has a volume of 1L.
Alternatively, the MH broth has a concentration of 5g/l, the brain heart infusion has a concentration of 40g/l, and the water has a volume of 1 l.
Optionally, the MH broth has a concentration of 27g/L, the brain heart infusion has a concentration of 5g/L, and the water has a volume of 1L.
The embodiment of the invention provides a preparation method of a culture medium composition, which comprises the following steps:
step one, cleaning an instrument used for weighing and dissolving the culture medium composition;
weighing the MH broth, the brain heart infusion and the water;
step three, dissolving the MH broth, the brain heart infusion and the water obtained by weighing to obtain a dissolved substance;
step four, filtering the dissolved substances to obtain a culture medium composition;
and fifthly, performing high-pressure damp-heat sterilization on the culture medium composition.
Optionally, the sterilized culture medium is subjected to sterility testing.
Optionally, filtering the dissolved material, including: the dissolved matter is subjected to primary filtration and then is subjected to secondary filtration, wherein the secondary filtration is carried out once, twice or three times.
Optionally, the first filtration uses a filter with a pore size of 0.3 m to 1 micron and the second filtration uses a filter with a pore size of 0.1 micron to 0.3 micron.
Optionally, the primary filtration uses a filter with a pore size of 0.4 microns and the secondary filtration uses a filter with a pore size of 0.22 microns.
Optionally, the sterilized culture medium is cultured at 35 ℃ for 48 hours, and then the sterility test is carried out.
Alternatively, a sterile-inspected acceptable medium is inoculated with clinically common bacteria to verify the culture capacity of the medium.
The use of the above-described culture medium in bacterial resistance counting.
The Chinese meaning of MH (Mueller-Hinton Broth, MH) Broth is hydrolyzed casein medium.
The technical scheme of the invention has the characteristics of accelerating the growth of microorganisms, reducing the background and reducing the occurrence of bacterial agglomeration growth, and the culture medium is applied to the counting of bacteria by a resistance method, so that the function of rapidly culturing the bacteria is achieved on one hand, and the requirements of reducing the background, namely reducing particulate matters, required by the resistance counting are met on the other hand. During the growth process of bacteria, the cocci have the phenomenon of agglomeration and aggregation growth, and the phenomenon achieves the aim of dispersing and growing the bacteria as much as possible through the brain heart immersion liquid in the technical scheme of the invention.
The technical scheme of the invention has good stability, can keep the stability of the granular materials within one year after the preparation is finished, and can ensure the culture capacity of bacteria; the content of the particulate matters is low, and the requirement of resistance counting can be met; the process is mature and the cost is low.
Description of{ Medium }
The present application discloses a medium that can be used for bacterial culture for bacterial enumeration. For example, the medium may be used for bacterial cultivation prior to counting results of a method and/or apparatus for bacterial counting to better achieve a method and/or apparatus for bacterial counting. For another example, the culture medium can be used as an in vitro enrichment culture reagent to provide suitable nutrients and growth environments for microbial growth and to accelerate the growth rate of microorganisms.
In some embodiments, the culture medium applied to bacterial counting may be applied in a variety of bacterial counting methods, such as counter assays (also may be referred to as microscopic counting methods), electronic counter counting methods (also may be referred to as resistance counting methods), living cell counting methods, gravimetric methods, and the like.
In some embodiments, a solvent may refer to a liquid used to dissolve and/or dilute other components (e.g., bacteria, fixatives, additives, etc.) in the composition. Exemplary solvents may include, but are not limited to, water, dimethyl sulfoxide, trifluoroacetic acid, methanol, ethanol, dimethylformamide, acetonitrile, acetic acid, acetone, pyridine, dioxane, chloroform, isopropanol, tetramethyl ethylenediamine, triethylamine, methyl formate, tributylamine, methyl ethyl ketone, ethyl acetate, trioctylamine, dimethyl carbonate, tetrahydrofuran, propanol, n-butanol, methylene chloride, benzene, diethyl ether, isopropyl ether, n-butyl ether, trichloroethylene, diphenyl ether, methylene chloride, toluene, carbon tetrachloride, carbon disulfide, cyclohexane, hexane, petroleum ether, and the like, or any combination thereof. Preferably, the solvent may be pure water. In some embodiments, the solvent may comprise 40% -75% of the total volume of the composition. More preferably, the solvent may comprise 41% to 70% of the total volume of the composition. More preferably, the solvent may comprise 42% to 65% of the total volume of the composition. More preferably, the solvent may comprise 43% to 60% of the total volume of the composition. More preferably, the solvent may comprise 44% to 59% of the total volume of the composition. More preferably, the solvent may comprise 45% -58% of the total volume of the composition. More preferably, the solvent may comprise from 46% to 57% of the total volume of the composition. More preferably, the solvent may comprise 47% to 56% of the total volume of the composition. More preferably, the solvent may comprise 48% to 54% of the total volume of the composition. More preferably, the solvent may comprise 54% of the total volume of the composition. More preferably, the solvent may comprise 53% of the total volume of the composition. More preferably, the solvent may comprise 52% of the total volume of the composition. More preferably, the solvent may comprise 51% of the total volume of the composition. More preferably, the solvent may comprise 50% of the total volume of the composition. More preferably, the solvent may comprise 49% of the total volume of the composition. Particularly preferably, the solvent may comprise 48% of the total volume of the composition.
Bacteria may be the final component of the composition that is counted. When the composition is used to verify a bacteria counting device or method, the value detected is the number of bacteria contained in the composition. In some embodiments, the bacteria may be selected from one or any combination of bacillus, coccus, helicobacter, and the like. Preferably, the bacteria may be selected from one or any combination of bacillus or coccus. Exemplary bacilli may include, but are not limited to, enterobacter aerogenes, enterobacter cloacae, enterobacter agglomerans, escherichia coli, escherichia friesen, escherichia helman, serratia marcescens, serratia rubra, klebsiella pneumoniae, proteus, providencia, salmonella, enterobacter trifoliatus, algobacter americanus, butje's bacteria, sildencia, shigella, acinetobacter, bifidobacterium, flavobacterium, bacillus, and the like, or any combination thereof. Exemplary cocci may include, but are not limited to, staphylococcus aureus, staphylococcus epidermidis, staphylococcus hemolyticus, staphylococcus intermedia, staphylococcus capitis, staphylococcus saccharolyticus, micrococcus, enterococcus chromeli, streptococcus bovis, streptococcus blood, streptococcus intermedia, streptococcus pyogenes, streptococcus beta hemolyticus, streptococcus angina, streptococcus twins, streptococcus pneumoniae, streptococcus turquoise, neisseria, and the like, or any combination thereof. Preferably, the bacteria may be selected from one of escherichia coli, staphylococcus aureus, or a combination thereof. Commercially available standard strains can be used in the compositions disclosed herein. By way of example only, the bacteria may be selected from the ATCC25922 standard strain of Escherichia coli, and/or the ATCC 29213 standard strain of Staphylococcus aureus. In some embodiments, the total concentration of bacteria may be 105-109/ml. More preferably, the total concentration of bacteria may be 106-108 bacteria/ml. More preferably, the total concentration of bacteria may be 106 bacteria/ml. More preferably, the total concentration of bacteria may be 107 bacteria/ml. Particularly preferably, the total concentration of the bacteria may be 108 bacteria/ml.
The additive may be a substance for adjusting the pH so that the components of the composition maintain their intact properties during storage. In some embodiments, the additive may be selected from inorganic salts including, but not limited to, sodium phosphate, potassium phosphate, calcium phosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium chloride, calcium chloride, potassium chloride, copper sulfate, iron sulfate, magnesium chloride, magnesium sulfate, sodium bicarbonate, zinc sulfate, and the like, or any combination thereof. Preferably, the additive may include one of potassium dihydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, sodium chloride, or any combination thereof. More preferably, the additive may include potassium dihydrogen phosphate, disodium hydrogen phosphate, and sodium chloride. In some embodiments, the amount of potassium dihydrogen phosphate included in the additive is 0.02% to 0.03% by weight of the total composition. More preferably, the amount of potassium dihydrogen phosphate included in the additive is 0.02% -0.027% of the total mass of the composition. More preferably, the amount of potassium dihydrogen phosphate included in the additive is 0.02% by weight of the total composition. More preferably, the potassium dihydrogen phosphate is included in the additive in an amount of 0.023% by mass of the total mass of the composition. More preferably, the amount of potassium dihydrogen phosphate included in the additive is 0.025% by weight of the total composition. Particularly preferably, the mass of potassium dihydrogen phosphate included in the additive is 0.027% of the total mass of the composition. The mass of the disodium hydrogen phosphate contained in the additive accounts for 0.1-0.2% of the total mass of the composition. More preferably, the disodium hydrogen phosphate is included in the additive in an amount of 0.11% to 0.18% by mass of the total mass of the composition. More preferably, the disodium hydrogen phosphate is included in the additive in an amount of 0.12% to 0.16% by mass of the total mass of the composition. More preferably, the disodium hydrogen phosphate is included in the additive in an amount of 0.13% to 0.15% by mass of the total mass of the composition. More preferably, the disodium hydrogen phosphate is included in the additive in an amount of 0.13% by mass of the total mass of the composition. More preferably, the disodium hydrogen phosphate is included in the additive in an amount of 0.14% by mass of the total mass of the composition. More preferably, the disodium hydrogen phosphate is included in the additive in an amount of 0.142% by mass of the total mass of the composition. The mass of sodium chloride contained in the additive accounts for 0.07% -0.1% of the total mass of the composition. More preferably, the sodium chloride is included in the additive in an amount of 0.08% -0.09% by mass of the total mass of the composition. More preferably, the sodium chloride is included in the additive in an amount of 0.08% by mass of the total mass of the composition. Particularly preferably, the sodium chloride is included in the additive in an amount of 0.09% by mass of the total mass of the composition.
It should be noted that the above description of the compositions, solvents, bacteria, fixatives, additives is for convenience of description only and is not intended to limit the application to the scope of the illustrated embodiments. It will be appreciated that any modifications and alterations to the parts are possible without departing from the principles of the present application, as will be apparent to those skilled in the art after having appreciated the principles. For example, the fixative may be replaced by a physical fixation method. Exemplary physical fixation methods may include paraffin infiltration methods, cryogenic freezing methods. Such variations are within the scope of the present application.
{ { preparation method }
One general method of preparing the media composition may include, for example, preparing a vessel for cleaning, weighing raw materials as required by the formulation, dissolving, filtering, autoclaving, and post-sterilization culturing. The preparation of the composition is described in further detail below.
Step one, cleaning preparation: and carrying out ultrasonic cleaning and pure water flushing on the container to reduce the content of particulate matters in the container.
In some embodiments, the cleaning means may include, but are not limited to, ultrasonic cleaning and pure water rinsing, as the exemplary cleaning means is to reduce the content of particulate matter in the container.
Secondly, weighing: weighing raw materials according to the requirements of the formula.
Third step, dissolving: and fully dissolving the weighed raw materials.
In some embodiments, the dissolution may include, but is not limited to, pure water dissolution, and the solvents used are described above and are not described herein.
Fourth, filtering: the presence of particulate material is reduced by first performing a primary filtration through a 0.4 μm filter and then performing a primary, secondary or tertiary filtration through a 0.22 μm filter.
In some embodiments, the filtering means may include, but is not limited to, direct filtration, membrane filtration, and the like, depending on the actual situation. Exemplary direct filtration means may include, but are not limited to, one or a combination of cartridge filtration, fine mesh filtration, and the like. Exemplary membrane filtration modes may include, but are not limited to, filtration modes of one or any combination of microporous filtration, ultrafiltration, reverse osmosis, and the like. Preferably, the dilution liquid can be filtered by a microporous filter membrane. In some embodiments, the microporous filter membrane may include one or any combination of cellulose acetate membrane, nitrocellulose membrane, polyamide membrane, polytetrafluoroethylene membrane, polyvinylidene fluoride membrane, surfactant-free cellulose acetate membrane, asymmetric polyethersulfone, fiberglass membrane, and the like. In some embodiments, the microporous filter membrane used may have a diameter of less than or equal to 12 μm (micrometers). More preferably, the microporous filter membrane used may have a diameter of less than or equal to 5 μm. More preferably, the microporous filter membrane used may have a diameter of less than or equal to 1 μm. More preferably, the microporous filter membrane used may have a diameter of less than or equal to 0.8 μm. More preferably, the microporous filter membrane used may have a diameter of less than or equal to 0.6 μm. More preferably, the microporous filter membrane used may have a diameter of less than or equal to 0.4 μm. More preferably, the microporous filter membrane may have a diameter of less than or equal to 0.3 μm. More preferably, the microporous filter membrane may have a diameter of less than or equal to 0.22 μm. More preferably, the microporous filter membrane may have a diameter of less than or equal to 0.12 μm.
Fifth, the filtered medium is subjected to high-pressure moist heat sterilization (inactivation).
In some embodiments, the inactivation may refer to an operation that disrupts the biological activity, reproductive ability, and pathogenicity of a microorganism (e.g., bacteria) to preserve its immunogenicity as much as possible. The means of inactivation may include physical inactivation methods and chemical inactivation methods. Exemplary physical inactivation may include one or any combination of high pressure wet heat inactivation, dry heat inactivation, ultrasonic inactivation, ultraviolet light inactivation, radiation irradiation inactivation, and the like. Exemplary chemical inactivation may include an inactivating agent of one or any combination of formaldehyde solution, alkylating agent, phenol, crystal violet, and the like. It will be appreciated that the effect of inactivation may be related to one or any combination of inactivation method, selection of the inactivating agent, inactivation temperature, inactivation time, pH, microorganism species, and the like. Preferably, the bacterial suspension may be inactivated by a method of high-pressure damp-heat bacterial inactivation. Parameters adopted in the high-pressure wet heat sterilization process can be adjusted according to actual conditions. For example, the parameters for the inactivation of high-pressure moist heat bacteria may be 121℃and 0.12MPa for 30min.
Sixth, the sterilized culture medium is cultured at 35 ℃ for 48 hours.
In some embodiments, the time of the culture may be adjusted according to the specific situation, and the above-mentioned "culturing of the sterilized culture medium at 35 ℃ for 48 hours" is only a preferred example, and the culture temperature and time may be adjusted according to the specific situation.
And seventh, bacteria after sterilization and culture are subjected to sterility inspection, and the culture capacity of a culture medium is checked by inoculating clinically common bacteria.
In some embodiments, the bacteria may be the last component to be counted in the composition. It should be noted that the bacteria added to the filtered dilution are live bacteria. The added bacteria may be selected from one or any combination of bacillus, coccus, helicobacter, etc. Preferably, the bacteria may be selected from the group consisting of escherichia coli and staphylococcus aureus. In some embodiments, methods of preparing a bacterial suspension may include, but are not limited to, turbidimeter or turbidimeter. The turbidimetric method can be a method for roughly judging the concentration or the number of bacteria in bacterial suspensions, and the concentration of the bacterial suspension to be detected is obtained by adjusting the turbidity between the bacterial suspensions to be detected to enable the turbidity of the bacterial suspension to be detected to be the same as or basically the same as the turbidity of a standard tube of the turbidimetric method. By way of example only, the turbidimetry may utilize sulfuric acid and barium chloride to formulate a mahalanobis standard tube number (e.g., 0.5 mahalanobis standard tube number, 1 mahalanobis standard tube number, 2 mahalanobis standard tube number, 3 mahalanobis standard tube number, 4 mahalanobis standard tube number), and the turbidity of the resulting barium sulfate precipitate may correspond to a bacterial concentration (e.g., 0.5 mahalanobis corresponding to a bacterial concentration of 1.5X108/ml, 1 mahalanobis corresponding to a bacterial concentration of 3X 108/ml, 2 mahalanobis corresponding to a bacterial concentration of 6X 108/ml, 3 mahalanobis corresponding to a bacterial concentration of 9X 108/ml, 4 mahalanobis corresponding to a bacterial concentration of 12X 108/ml). And judging the concentration of the bacterial suspension by comparing the turbidity of the bacterial suspension to be detected with that of a Maillard standard tube. As another example, the turbidimeter can directly display turbidity values of the units of mahogany by measuring the bacterial suspension to be measured, and can obtain the bacterial suspension concentration to be measured from a table of the relationship between international universal turbidimeter values (e.g., 0.5McF, 1McF, 2McF, 3McF, 4 McF) and the concentration of the bacterial suspension (e.g., 0.5McF corresponds to 1.5×108/ml, 1McF corresponds to 3×108/ml, 2McF corresponds to 6×108/ml, 3McF corresponds to 9×108/ml, and 4McF corresponds to 12×108/ml).
It should be noted that the above description of the process for preparing the medium composition is for illustration and description only, and does not limit the scope of applicability of the application. It will be apparent to those skilled in the art having the benefit of this disclosure that many modifications and changes to this process may be made without departing from the scope of the invention. Such modifications and variations are, however, still within the scope of the present application.
{ { use of Medium composition in bacterial enumeration method or apparatus }
In some embodiments, the culture medium composition may be used for bacterial culture for bacterial enumeration. For example, bacteria are grown rapidly to meet the background-reducing, i.e., particulate matter-reducing, requirements such as resistance counts.
In the first step, a suitable vessel is selected for bacterial culture.
In some embodiments, the bacteria are as described above and are not described in detail herein.
And secondly, adding the culture medium composition into the bacteria, and culturing the bacteria for less than or equal to 4 hours.
Third, the number of bacteria in the composition (composition containing bacteria and its medium) is measured by a bacteria counting method or apparatus, to obtain a measurement of the number of bacteria in the composition.
In some embodiments, the bacterial count method may include one or any combination of electronic counting, microscopy, living cell counting, cell weight method, and the like. The bacteria counting device may be a device for performing bacteria counting using the bacteria counting method as described above. The measured value of the bacterial concentration may be a value obtained by counting the bacteria in the composition in an existing step using the bacterial counting method or in an existing state using the bacterial counting apparatus. The measured value can be obtained by implementing all the steps of the bacterial counting or operating the bacterial counting device. Preferably, the bacterial count method is a resistance count method.
Possible beneficial effects of embodiments of the present application include, but are not limited to: the composition disclosed by the application can be used for the accelerated culture of bacteria in bacteria counting equipment or methods, and has the advantages of high accuracy, good stability and high speed. It should be noted that, the advantages that may be generated by different embodiments may be different, and in different embodiments, the advantages that may be generated may be any one or a combination of several of the above, or any other possible advantages that may be obtained.
Detailed Description
The principles and spirit of the present invention will be described below with reference to several exemplary embodiments. It should be understood that these embodiments are presented merely to enable those skilled in the art to better understand and practice the invention and are not intended to limit the scope of the invention in any way. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
The present application is further illustrated by the following specific examples.
EXAMPLE 1 preparation of the Medium composition
The preparation process is as follows:
firstly, placing the container into an ultrasonic cleaner for cleaning, and flushing with pure water for three times after the cleaning is finished.
And secondly, weighing MH broth and brain heart infusion according to a formula, and dissolving with water.
And thirdly, filtering the dissolved substances, namely, firstly, performing primary filtration through a 0.4 mu m filter membrane, and then performing primary, secondary or tertiary filtration through a 0.22 mu m filter membrane to reduce the existence of particulate matters, namely, performing filtration according to the filtration conditions of different proportions identified in the table.
Fourth, the filtered culture medium composition is subjected to high-pressure moist heat sterilization.
And fifthly, culturing the sterilized culture medium composition at 35 ℃ for 48 hours, performing sterility test on the sterilized culture medium composition, and inoculating the common bacteria to test the culture capacity of the culture medium. Culturing at 35℃for 48 hours is only an example, and the temperature may be between 30℃and 40℃for 36 hours to 72 hours.
The formulation of the medium composition is shown in Table 1.
Table 1.
| Main component | MH | Brain heart infusion | Pure water |
| A culture medium formula | 21g/L | 12g/L | 1L |
| B culture medium formula | 5g/L | 40g/L | 1L |
| C culture medium formula | 27g/L | 5g/L | 1L |
| D culture medium formula | 0.5g/L | 60g/L | 2L |
| E culture medium formula | 40g/L | 0.5g/L | 2L |
| F culture medium formula | 4g/L | 10g/L | 2L |
| G culture medium formula | 20g/L | 30g/L | 2L |
| H culture medium formula | 10g/L | 50g/L | 1L |
| I culture medium formula | 18g/L | 15g/L | 1L |
| J Medium formulation | 25g/L | 20g/L | 1L |
Example 2-preparation of the culture medium composition the preparation process was as follows:
firstly, placing the container into an ultrasonic cleaner for cleaning, and flushing with pure water for three times after the cleaning is finished.
In the second step, MH broth 21g/L (g/L) and brain heart infusion 12g/L were weighed and dissolved in 1L water.
In a third step, the solubilizate is filtered, first through a 0.4 μm (micrometer) filter, and then through a 0.22 μm filter for three times to reduce the presence of particulate material.
Fourth, the filtered culture medium composition is subjected to high-pressure moist heat sterilization.
And fifthly, culturing the sterilized culture medium composition for 48 hours at 35 ℃, performing sterility test on the sterilized culture medium composition, and inoculating clinical common bacteria to test the culture capacity of the culture medium for later use to obtain the culture medium composition A.
Example 3-preparation of the culture medium composition the preparation process was as follows:
and firstly, placing the container into an ultrasonic cleaner for cleaning, and flushing twice with pure water after the cleaning is finished.
In the second step, 5g/L (g/L) of MH broth and 40g/L of brain heart infusion were weighed and dissolved in 1L of water.
In a third step, the solubilizate is filtered, first through a 0.4 μm (micrometer) filter, and then through a 0.22 μm filter to reduce the presence of particulate material.
Fourth, the filtered culture medium composition is subjected to high-pressure moist heat sterilization.
And fifthly, culturing the sterilized culture medium composition for 48 hours at 35 ℃, performing sterility test on the sterilized culture medium composition, and inoculating clinical common bacteria to test the culture capacity of the culture medium for later use to obtain the culture medium composition B.
Example 4-preparation of the culture medium composition the preparation process was as follows:
and firstly, placing the container into an ultrasonic cleaner for cleaning, and flushing twice with pure water after the cleaning is finished.
In the second step, 27g/L (g/L) of MH broth and 5g/L of brain heart infusion were weighed and dissolved in 1L of water.
In a third step, the solubilizate is filtered, first through a 0.4 μm (micrometer) filter, and then through a 0.22 μm filter to reduce the presence of particulate material.
Fourth, the filtered culture medium composition is subjected to high-pressure moist heat sterilization.
And fifthly, culturing the sterilized culture medium composition at 35 ℃ for 48 hours, performing sterility test on the sterilized culture medium composition, and inoculating clinical common bacteria to test the culture capacity of the culture medium for later use to obtain the composition of the culture medium composition C.
EXAMPLE 5 preparation of the Medium composition
The preparation procedure was similar to that of examples 2 to 4, and a D medium composition, an E medium composition, an F medium composition, a G medium composition, an H medium composition, an I medium composition, and a J medium composition were prepared according to the formulation composition of the medium composition of example 1.
Example 6-optimal Medium composition formulation screening purposes: through preparing the culture medium composition, the aim of ensuring the rapid growth of clinically common bacteria and achieving the bacterial count in as short a time (1-4) as possible is selected, namely the aim of rapidly counting the bacterial count by resistance is fulfilled.
The experimental steps are as follows:
placing the container into an ultrasonic cleaner for cleaning, and flushing with pure water for three times after the cleaning is finished;
weighing and dissolving according to the formula requirement;
the dissolved reagent was subjected to primary filtration (0.4 μm filter), and after completion of filtration, the reagent was subjected to filtration with 0.22 μm filter three times, and the results were recorded by resistance counting, or the test was performed directly using the medium compositions of examples 2 to 4.
The time required for inoculating 50cfu/ml of Escherichia coli (ATCC 25922), pseudomonas aeruginosa (ATCC 27853), staphylococcus aureus (ATCC 25923), enterococcus faecalis (ATCC 29212) and Streptococcus pneumoniae (ATCC 49619) to reach a value of about 200 was recorded, and the test results are shown in Table 2.
TABLE 2
Time/h is time/hour experimental result analysis:
according to five standard strains in the test result, bacterial growth can be satisfied within the range of the formula for 4 hours to reach a resistance count value of about 200, wherein different proportions of MH and brain heart infusion can lead to inconsistent growth time when the resistance count of various bacteria reaches 200, wherein the formula of the culture medium A is the optimal proportion, and the time required for the growth of the five standard strains is the shortest, namely the optimal value.
EXAMPLE 7 screening of Process for preparation of Medium composition
The purpose is as follows: by preparing the culture medium composition, the optimal filtration mode is selected to remove the particulate matter so as to ensure that the culture medium can meet the requirements of bacterial culture resistance count.
The experimental steps are as follows:
1. placing the container into an ultrasonic cleaner for cleaning, and flushing with pure water for three times after the cleaning is finished;
2. weighing and dissolving according to the requirements of the culture medium formula A;
3. the dissolved reagent was subjected to primary filtration (0.4 μm filter), and after completion of filtration, the reagent was filtered once, twice and three times with 0.22 μm filter, and the number of particulate matters was counted by electric resistance, and the results are shown in Table 3;
TABLE 3 Table 3
Analysis of experimental results: according to the test result, the filter membrane is analyzed to remove 43% of the particulate matters once, 92.6% of the particulate matters are removed twice, 98.6% of the particulate matters are removed three times, the requirement of reducing the bacterial count background is completely met, the dissolved reagent is subjected to primary filtration by a 0.4 mu m filter membrane, the particulate matters removing effect of the culture medium composition after the filtration is finished by the 0.22 mu m filter membrane for three times is optimal, the requirement of resistance bacteria count on the minimum of the particulate matters can be met, and unexpected effects are generated.
While the basic concepts have been described above, it will be apparent to those skilled in the art that the foregoing detailed disclosure is by way of example only and is not intended to be limiting. Although not explicitly described herein, various modifications, improvements, and adaptations of the present application may occur to one skilled in the art. Such modifications, improvements, and modifications are intended to be suggested within this application, and are therefore within the spirit and scope of the exemplary embodiments of this application.
Meanwhile, the present application uses specific words to describe embodiments of the present application. Reference to "one embodiment," "an embodiment," and/or "some embodiments" means that a particular feature, structure, or characteristic is associated with at least one embodiment of the present application. Thus, it should be emphasized and should be appreciated that two or more references to "an embodiment" or "one embodiment" or "an alternative embodiment" in various positions in this specification are not necessarily referring to the same embodiment. Furthermore, certain features, structures, or characteristics of one or more embodiments of the present application may be combined as suitable.
Furthermore, those skilled in the art will appreciate that the various aspects of the invention are illustrated and described in the context of a number of patentable categories or circumstances, including any novel and useful procedures, machines, products, or materials, or any novel and useful modifications thereof.
Furthermore, the order in which the elements and sequences are presented, the use of numerical letters, or other designations are used in the application and are not intended to limit the order in which the processes and methods of the application are performed unless explicitly recited in the claims. While certain presently useful inventive embodiments have been discussed in the foregoing disclosure, by way of various examples, it is to be understood that such details are merely illustrative and that the appended claims are not limited to the disclosed embodiments, but, on the contrary, are intended to cover all modifications and equivalent arrangements included within the spirit and scope of the embodiments of the present application.
Likewise, it should be noted that in order to simplify the presentation disclosed herein and thereby aid in understanding one or more inventive embodiments, various features are sometimes grouped together in a single embodiment, figure, or description thereof. This method of disclosure, however, is not intended to imply that more features than are presented in the claims are required for the subject application. Indeed, less than all of the features of a single embodiment disclosed above.
In some embodiments, numbers describing the components, number of attributes are used, it being understood that such numbers being used in the description of embodiments are modified in some examples by the modifier "about," approximately, "or" substantially. Unless otherwise indicated, "about," "approximately," or "substantially" indicate that the number allows for a 20% variation. Accordingly, in some embodiments, numerical parameters set forth in the specification and claims are approximations that may vary depending upon the desired properties sought to be obtained by the individual embodiments. In some embodiments, the numerical parameters should take into account the specified significant digits and employ a method for preserving the general number of digits. Although the numerical ranges and parameters set forth herein are approximations that may be employed in some embodiments to confirm the breadth of the range, in particular embodiments, the setting of such numerical values is as precise as possible.
Each patent, patent application publication, and other material, such as articles, books, specifications, publications, documents, etc., cited in this application is hereby incorporated by reference in its entirety. Except for application history documents that are inconsistent or conflicting with the present application, documents that are currently or later attached to this application for which the broadest scope of the claims to the present application is limited. It is noted that the descriptions, definitions, and/or terms used in the subject matter of this application are subject to such descriptions, definitions, and/or terms if they are inconsistent or conflicting with such descriptions, definitions, and/or terms.
Finally, it should be understood that the embodiments described herein are merely illustrative of the principles of the embodiments of the present application. Other variations are also possible within the scope of this application. Thus, by way of example, and not limitation, alternative configurations of embodiments of the present application may be considered in keeping with the teachings of the present application. Accordingly, embodiments of the present application are not limited to only the embodiments explicitly described and depicted herein.
Claims (9)
1. Use of a culture medium composition for bacterial resistance counting, characterized in that the composition of the culture medium composition is: MH broth at a concentration of 0.5 g/l to 40g/l, brain heart infusion at a concentration of 0.5 g/l to 60 g/l, and water.
2. Use of a culture medium composition according to claim 1 for bacterial resistance counting, characterized in that the composition of the culture medium composition is: MH broth at a concentration of 5g/l to 27g/l, brain heart infusion at a concentration of 5g/l to 40g/l, and water.
3. Use of a culture medium composition according to claim 2 for bacterial resistance counting, characterized in that the concentration of MH broth is 21g/l, the concentration of brain heart infusion is 12g/l, and the volume of water is 1 l.
4. Use of a culture medium composition according to claim 2 for bacterial resistance counting, characterized in that the concentration of MH broth is 5g/l, the concentration of brain heart infusion is 40g/l, and the volume of water is 1 l.
5. Use of a culture medium composition according to claim 2 for bacterial resistance counting, characterized in that the concentration of MH broth is 27g/l, the concentration of brain heart infusion is 5g/l, and the volume of water is 1 l.
6. Use of a culture medium composition according to any one of claims 1 to 5 for bacterial resistance counting, characterized in that the preparation method of the culture medium composition comprises:
cleaning an instrument used for weighing and dissolving the culture medium composition;
weighing the MH broth, the brain heart infusion and the water;
dissolving the weighed MH broth, brain heart infusion and water to obtain a dissolved substance;
filtering the dissolved matter to obtain a culture medium composition;
the culture medium composition is sterilized.
7. The use of the culture medium composition according to claim 6 for bacterial resistance counting, wherein filtering the lysate comprises: the lysate is first filtered and then filtered again, wherein the number of times of the filtering again is one, two or three.
8. The use of the culture medium composition according to claim 7, wherein the primary filtration uses a filter pore size of 0.3 to 1 micron and the secondary filtration uses a filter pore size of 0.1 to 0.3 micron.
9. The use of the culture medium composition according to claim 8, wherein the primary filtration uses a filter pore size of 0.4 microns and the secondary filtration uses a filter pore size of 0.22 microns.
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| US4103229A (en) * | 1977-01-28 | 1978-07-25 | The University Of Virginia | Continuous-flow, resistive-particle counting apparatus |
| WO2010035458A1 (en) * | 2008-09-29 | 2010-04-01 | 三愛石油株式会社 | Culture medium for determining total viable cell count |
| CN102459571A (en) * | 2009-06-04 | 2012-05-16 | 香港中文大学 | Anti-inflammatory bacteria |
| CN105749260A (en) * | 2016-05-11 | 2016-07-13 | 山东司邦得制药有限公司 | Lysozyme hydrochloride vaginal tablets, and preparation method and application thereof |
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