資源描述:
《美國藥典無菌檢查法》由會員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫。
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