美國藥典無菌檢查法

美國藥典無菌檢查法

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時間:2019-11-28

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1、Accessedfrom64.40.16.20bykows3buckonThuAug1400:50:28EDT2014USP37MicrobiologicalTests/á71?SterilityTests71detectingMycoplasmas.Testtheeffectivenessofthecellsto5.AddasuitableDNAstainandallowstandingforasuit-beusedbyapplyingtheprocedureshownbelowandinocu-abletime(bisbenzimideworkings

2、olutionandastand-latingnotmorethan100cfuorccumicroorganismsofsuit-ingtimeof10minutesaresuitable).ablereferencestrainsofM.hyorhinisandM.orale.Thecells6.RemovethestainandrinsethemonolayerwithPurifiedaresuitableifbothreferencestrainsaredetected.Theindi-Water.catorcellsmustbesubcultur

3、edwithoutanantibioticbefore7.Mounteachcoverslip,whereapplicable(amixtureofuseinthetest.equalvolumesofglycerolandPhosphate-CitrateBufferSolutionpH5.5issuitableformounting).ExaminebyTestMethodfluorescence(forbisbenzimidestaina330nm/380nmexcitationfilterandanLP440nmbarrierfilteraresu

4、it-NOTE—Thefollowingisprovidedforinformation.able)at400×magnificationorgreater.8.Comparethemicroscopicappearanceofthetestcul-SOLUTIONStureswiththatofthenegativeandpositivecontrols,examiningforextranuclearfluorescence.MycoplasmasproducepinpointsorfilamentsovertheindicatorcellPhosph

5、ateBufferedSaline—cytoplasm.Theymayalsoproducepinpointsandfila-2.0MMonobasicPotassiumPhosphate—Dissolve13.61gmentsintheintercellularspaces.Multiplemicroscopicofanhydrousmonobasicpotassiumphosphatein50mLoffieldsareexaminedaccordingtotheprotocolestablish-water.edduringvalidation.2.0

6、MDibasicPotassiumPhosphate—Dissolve17.42gofGeneralChaptersanhydrousdibasicpotassiumphosphatein50mLofwater.PhosphateBufferedSalineSolution(pH7.4)—Combine3.6InterpretationofResultsmLof2.0MMonobasicPotassiumPhosphate,16.4mLof2.0MDibasicPotassiumPhosphate,8gofsodiumchloride,andTheprod

7、ucttobeexaminedcomplieswiththetestifflu-1Lofwater.Mixthoroughly.AdjustthepHifnecessary.orescencetypicalofMycoplasmasisnotpresent.ThetestisBisbenzimideStockSolution—Dissolve5mgofbisben-invalidifthepositivecontrolsdonotshowfluorescencetyp-zimideinwater,anddilutewiththesamesolventto1

8、00icalofMycoplasmas.Thetestisinva

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