美国药典-USP36微生物限度检查

USP361117优良微生物检测规范（中英文1/2）2013-08-0915:30:46|分类：USP|举报|字号订阅1117MICROBIOLOGICALBESTLABORATORYPRACTICES优良微生物检测规范INTRODUCTION介绍Goodlaboratorypracticesinamicrobiologylaboratoryconsistofactivitiesthatdependonseveralprinciples:aseptictechnique,controlofmedia,controlofteststrains,operationandcontrolofequipment,diligentrecordingandevaluationofdata,andtrainingofthelaboratorystaff.Becauseoftheinherentriskofvariabilityinmicrobiologydata,reliabilityandreproducibilityaredependentontheuseofacceptedmethodsandadherencetogoodlaboratorypractices.优良微生物检测规范由一些活动组成，这些活动依赖于几个基本要素：无菌技术、培养基控制、检测用菌株控制、设备操作和控制、完善的记录和数据评估、化验室员工的培训。由于微生物数据具有天生的不确定性，数据的可靠性和重复性取决于是否使用被接受的方法，以及是否严格遵守化验室规范。MEDIAPREPARATIONANDQUALITYCONTROL培养基制备和质量控制MediaPreparation培养基制备Culturemediaarethebasisformostmicrobiologicaltests.Safeguardingthequalityofthemediaisthereforecriticaltothesuccessofthemicrobiologylaboratory.Mediapreparation,properstorage,andqualitycontroltestingcanensureaconsistentsupplyofhigh-qualitymedia.培养基是大多数微生物测试的基础。保证培养基的质量因而成为微生物实验室成功的关键。培养基的制备、合适的存贮和质量控制检测可以保证持续高质量培养基供应。Itisimportanttochoosethecorrectmediaorcomponentsinmakingmediabasedontheuseofacceptedsourcesorreferencesforformulas.Themanufacturer'sformulaandinstructionsforpreparationroutinelyaccompanydehydratedmediaandready-mademedia.Becausedifferentmediatypesmayhavedifferentpreparationrequirements(e.g.,heating,additives,andpHadjustment),itisimportanttofollowtheseinstructionstoensurepreparationofacceptablemediaquality.Acertificateofanalysisdescribingexpirationdatingandrecommendedstorageconditionsaccompaniesready-mademedia,aswellasthequalitycontrolorganismsusedingrowth-promotionandselectivitytestingofthatmedia.在制备培养基过程中使用已接受的来源或配方标准，选择正确的培养基或成份是非常重要的。一般生产商在供应培养基干粉和配制好的培养基时，其配方和配制指示都会随货发送。由于不同的培养基类型可能有不同的配制要求（例如，加热、添加剂，和pH值调节），重要的一点是需要遵守其提供的配制指示以保证配制出的培养基质量。如果是已制备好的培养基碟/瓶，随货会收到指明有效期和推荐的存贮条件的分析报告，以及促生产试验用微生物种类，和该培养基的选择性试验用微生物种类。Wateristheuniversaldiluentformicrobiologicalmedia.PurifiedWaterismostoftenusedformediapreparation,butincertaincasestheuseofdeionizedordistilledwatermaybeappropriate.Wateroflesserqualityshouldnotbeusedformicrobiologicalmediapreparation.Thevolumeofthewaterusedshouldberecorded.水是通用的微生物培养基稀释剂。纯化水常用于培养基制备，但在某些情况下，也可以使用去离子水或蒸馏水。更低品质的水不应用于微生物培养基制备。水的用量应记录。Consistentpreparationofmediarequiresaccurateweighingofdehydratedmediaormediaconstituents.Acalibratedbalancewiththeappropriateweightrangefortheingredientsshouldbeused(SeeWeighingonanAnalyticalBalance1251).Cleanweighingcontainersandtools(suchasspatulas)shouldbeusedtopreventforeignsubstancesfromenteringtheformulation.Theweightofthecomponentsshouldberecorded.要保持培养基配制的一致性，需要对培养基干粉或培养基组份进行准确称量。应使用在所需称量范围内经过校正的天平（见1251分析天平称量）。应对称重容器和工具（例如料勺）进行清洁以保证无异物进入所配物料。各成份的重量应进行记录。Dehydratedmediashouldbethoroughlydissolvedinwaterbeforedispensingandsterilization.Ifheatingisnecessarytohelpdissolvethemedia,careshouldbetakennottooverheatmedia,becauseallculturemedia,toagreaterorlesserextent,areheat-sensitive.Equipmentusedinthepreparationofmediashouldbeappropriatetoallowforcontrolledheating,constantagitation,andmixingofthemedia.培养基干粉应在水中完全溶解，然后进行配制和灭菌。如果需要加热助溶，要注意不能过热，因为所有的培养基，或多或少，都是对热敏感的。用于培养基配制的设备应适当，以便控制加热、持续搅拌和培养基混合。Darkeningofmedia(Maillard-typereactionornonenzymaticbrowning)isageneralindicationofoverheating.Whenaddingrequiredsupplementstomedia,adequatemixingofthemediumafteraddingthesupplementshouldbeperformed.培养基变黑（美拉德类型反应或非酶褐变）一般说明过热。在向培养基中加入所需要的补充成分时，在加入后需要进行充分搅拌混合。Preparationofmediainpoorlycleanedglasswarecanallowinhibitorysubstancestoenterthemedia.在清洁不彻底的玻璃器皿中配制培养基会使得抑制性物质带入培养基。Inhibitorysubstancescancomefromdetergentresidueaftercleaningglasswareorfrompriormaterialsusedintheglassware.Besurethatthecleaningprocessremovesdebrisandforeignmatter,andthatthedetergentisthoroughlyrinsedoutwithPurifiedWater.SeeCleaningGlassApparatus1051foradditionalguidance.抑制性物质可能来自于玻璃器皿中的清洁剂残留，或来自于玻璃器皿中上次所盛装的物料。要保证清洗程序可以去除残渣和外来物质，并且清洁剂可以被纯化水彻底冲洗掉。参见1051玻璃容器的清洁。Sterilizationofmediashouldbeperformedwithintheparametersprovidedbythemanufacturerorvalidatedbytheuser.Commerciallypreparedmediashouldprovidedocumentationofthesterilizationmethodused.培养基灭菌应在生产商提供的参数范围，或用户验证的参数范围内实施。商业制备好的培养基应随货有所用的灭菌方法。Autoclavingbymoistheatisthepreferredsterilizationtechnique,exceptininstanceswhenboilingisrequiredinordertoavoiddeteriorationofheat-labilecomponentsofthemedia.Sterilizationbyfiltrationmayalsobeappropriateforsomeformulations.湿热灭菌是较好的灭菌技术，除非需要煮沸以避免培养基中不耐热成分被破坏。有些配方可能适用过滤灭菌。Theeffectsofthesterilizationmethodandconditionsonthemediashouldbevalidatedbysterilityandgrowth-promotiontestingofthemedia.Inaddition,ifsterilizedbymoistheat,theautoclavecycleshouldbevalidatedtoensureproperheatdistributionforselectedloadsandvolumes.Typically,manufacturersrecommendusinganautoclavecycleof121for15minutesusingavalidatedautoclave.Theseconditionsapplytotimeattemperatureofthemedia.Ascontainersizeandtheloadconfigurationoftheautoclavewillinfluencetherateofheating,longercyclesmayberequiredforlargerloads.However,thesterilizationtimewillbedependentonthemediavolumeandautoclaveload.Sterilizationcyclesinwhichtheau