USP361117优良微生物检测规范中英文版

1、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,operationandcontrolofequipm

2、ent,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

3、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,properstora

4、ge,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.Themanufacturersformulaandinstructions

5、forpreparationroutinelyaccompanydehydratedmediaandready-mademedia.Becausedifferentmediatypesmayhavedifferentpreparationrequirements(e.g.,heating,additives,andpHadjustment),itisimportanttofollowtheseinstructionstoensurepreparationofacceptablemediaquality.Acertificateofanalysisdescribingexpirationdati

6、ngandrecommendedstorageconditionsaccompaniesready-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

7、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.Wateroflesserqualityshouldnotbeusedformicrobiologicalmediapreparat

8、ion.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(SeeWeighingona

9、nAnalyticalBalance1251).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。Dehydratedmed

10、iashouldbethoroughlydissolvedinwaterbeforedispensingandsterilization.Ifheatingisnecessarytohelpdissolvethemedia,careshouldbetakennottooverheatmedia,becauseallculturemedia,toagreaterorlesserextent,areheat-sensitive.Equipmentusedinthepreparationofmediashouldbeappropriatetoallowforcontrolledheating,con

11、stantagitation,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,adequatemixingofthemediumafteraddingth

12、esupplementshouldbeperformed.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。Inhibitorysubstancescancomefromdetergentresidueaftercleaningglasswareorfrompriormaterialsusedin

13、theglassware.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。Sterilizationofmediashouldbepe

14、rformedwithintheparametersprovidedbythemanufacturerorvalidatedbytheuser.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,exceptininstanceswhenboilingisrequir

15、edinordertoavoiddeteriorationofheat-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.In

16、addition,ifsterilizedbymoistheat,theautoclavecycleshouldbevalidatedtoensureproperheatdistributionforselectedloadsandvolumes.Typically,manufacturersrecommendusinganautoclavecycleof121for15minutesusingavalidatedautoclave.Theseconditionsapplytotimeattemperatureofthemedia.Ascontainersizeandtheloadconfig

17、urationoftheautoclavewillinfluencetherateofheating,longercyclesmayberequiredforlargerloads.However,thesterilizationtimewillbedependentonthemediavolumeandautoclaveload.Sterilizationcyclesinwhichtheautoclaveisslowtocomeuptotemperaturemayresultinoverheatingofthemedia.Therefore,caremustbetakentovalidate

18、asterilizationcycle,balancingtheneedforsterilemediaagainstthetendencyofthemediatodegradeunderexcessiveheating.Storageofthemediaintheautoclaveaftertheliquidcycleiscompletedisnotrecommendedaftercooling,asitmaydamagethemedia.Improperheatingorsterilizingconditionsforcommerciallypreparedorinternallyprepa

19、redmediamayresultinadifferenceincolorchange,lossofclarity,alteredgelstrength,orpHdriftfromthemanufacturersrecommendedrange,aswellasreducedgrowth-promotionactivityand/orselectivity.灭菌方法和条件对培养基的影响应经过无菌验证和培养基促生长试验确认。另外，如果采用湿热灭菌，灭菌周期应进行验主下以保证在所选的负载和体积下的热分布。生产商一般会推荐采用121C15分钟作为灭菌条件，培养基灭菌即采用此条件。由于容器尺寸和灭菌器

20、的负载参数会影响加热速度，如果负载较大时，应延长灭菌时间。当然，灭菌时间还是取决于培养基体积和灭菌负载。升温较慢的灭菌条件可能会导致培养基过热，因此需要特别注意灭菌周期的验证，在培养期灭菌要求和培养基在过热条件下分解中寻求平衡点。在灭菌结束冷却后，不建议将培养基留在灭菌器中存贮，因为可能会对培养基造成损坏。不适当的加热或灭菌条件对于商业制备的培养基或公司自制的培养基可能会导致颜色变化差异、澄清度差、胶强度改变、或pH值超出生产商指定范围、以及促生产试验表示出活性下降和/或具有选择性。ThepHofeachbatchofmediumshouldbeconfirmedafterithascoole

21、dtoroomtemperature(20-25)byasepticallywithdrawingasamplefortesting.RefrigeratedpurchasedmediashouldbeallowedtowarmuptoambientroomtemperatureifitistobecheckedforpHconfirmation.AflatpHprobeisrecommendedforagarsurfaces,andanimmersionprobeisrecommendedforliquids.SeepH791forguidancewithpHmeasurementandin

22、strumentcalibration.ThepHofmediashouldbeinarangeof0.2ofthevalueindicatedbythemanufacturer,unlessawiderrangeisacceptablebythevalidatedmethod.每个批次培养基在冷却到室温后（20-25C），应采用无菌方式取样检测pH值；冷冻的采购来的培养基应在升温至室温后检测pH值。建议对培养基平面采用扁平的pH探头，液体培养基则采用浸入式探头。pH值测量和仪器校正指南见pH791.除非有验证过的方法可以接受一个更宽的范围，否则培养基的pH值应在生产商指示的pH值0.2之内。

23、Preparedmediashouldbecheckedbyappropriateinspectionofplatesandtubesforthefollowing:培养基制备好后，应对碟或管进行以下检查Crackedcontainersorlids容器或盖裂开Unequalfillingofcontainers容器填充不匀Dehydrationresultingincracksordimpledsurfacesonsolidmedium脱水导致固体培养基裂开或凸陷Hemolysis溶血Excessivedarkeningorcolorchange太黑或颜色变化Crystalformation

24、frompossiblefreezing可能因为冷冻引起的结晶Excessivenumberofbubbles过量气泡Microbialcontamination微生物污染Statusofredoxindicators(ifappropriate)氧化还原指示剂状态(适用时)Lotnumberandexpirationdatecheckedandrecorded检查批号和有效期并记录Sterilityofthemedia培养基灭菌Cleanlinessofplates(lidshouldnotsticktodish)培养基用碟清洁(盖不应该粘在碟上)MediaStorage培养基存贮Itisp

25、rudenttoconsiderhowthemanufacturerorsuppliertransportsandstoresmediabeforedistributiontotheenduser.Manufacturersofmediashouldusetransportandstorageconditionsthatminimizethelossofmoisture,controlthetemperature,preventmicrobialcontamination,andprovidemechanicalprotectiontothepreparedmedia.培养基的生产商或供应商在

26、培养基销售给最终用户前的运输和存贮方式是需要慎重考虑的。培养基生产商所采用的运输和贮存条件应最大程度降低水份损失、保证温度的控制、防止微生物污染，提供对已制备培养基的物理保护。Mediashouldbelabeledproperlywithbatchorlotnumbers,preparationandexpirationdates,andmediaidentification.Mediashouldbestoredaccordingtothemanufacturersinstructions.Mediapreparedinhouseshouldbestoredundervalidatedco

27、nditions.Donotstoreagaratorbelow0C,asfreezingcoulddamagethegelstructure.Protectstoredmediafromexposuretolightandexcessivetemperature.Beforeprolongedstorage,agarplatesshouldbeplacedintoasealedpackageorcontainertoretardmoistureloss.培养基应标识批号，制备时间和有效期，以及培养基识别信息。培养基应根据生产商指示进行存贮。公司自己制备的培养基应在经过验证条件下存贮，不要将培

28、养基存贮等于或低于oc条件下，因为结冻可能会对胶质结构造成损伤。存贮期间培养基应避光，避免超温。如果要延长存贮时间，培养基碟应放在密封的包装或容器中以减少水分损失。Remeltingofanoriginalcontainerofsolidmediashouldbeperformedonlyoncetoavoidmediawhosequalityiscompromisedbyoverheatingorpotentialcontamination.Itisrecommendedthatremeltingbeperformedinaheatedwaterbathorbyusingfree-flowi

29、ngsteam.Theuseofmicrowaveovensandheatingplatesiscommon,butcareshouldbetakentoavoiddamagingmediabyoverheatingandtoavoidthepotentialinjurytolaboratorypersonnelfromglassbreakageandburns.Themoltenagarmediumshouldbeheldinamonitoredwaterbathatatemperatureof45to5ofornotmorethan8hours.固体培养基只能解冻一次，以避免培养基由于过热

30、或潜在污染造成质量问题。建议在热水浴中或使用自由流动蒸汽中解冻培养基。通常会使用微波炉和加热盘，但需要注意避免因为过热造成培养基损伤，避免因玻璃仪器破裂和燃烧引起化验室人员受伤。熔融的培养基应在40-45C水浴中不超过8小时。Cautionshouldbetakenwhenpouringthemediafromacontainerimmersedinawaterbathtopreventwaterfromthebathcomminglingwiththepouredsterilemedia.Wipingtheexteriorofthecontainerdrybeforepouringmaybe

31、advisable.将培养基从浸在水浴中的容器中倒出时要特别注意，避免水浴用水混入倒出的无菌培养基中。最好在将容器从水浴中取出后、倾倒前，将容器外表面擦干。Disposalofusedculturedmedia(aswellasexpiredmedia)shouldfollowlocalbiologicalhazardsafetyprocedures.使用过的培养基处理(和过期的培养基)需要按照微生物危险控制程序处理。QualityControlTesting质量控制检测Althoughgrowthmediacanbepreparedinalaboratoryfromindividualcom

32、ponents,manylaboratories,foreaseofuse,usedehydratedmediaorpurchasecommerciallypreparedmediainplasticplatesorglasscontainers.Manufacturersofmediaattempttostandardizerawmaterialsfrombiologicalsources,butmustconstantlydealwithunavoidabledifferencesinrawmaterialsobtainedfromnaturalsources,andtherefore,l

33、ot-to-lotvariabilityofmediamustbeconsidered.Inaddition,theperformanceofmediapreparedinalaboratoryorbyamanufacturerishighlydependentonpreparationandstorageconditions.虽然生产用培养基可以采用各组份配制而成，但许多化验室为了方便，采用培养基干粉或采购商业制备的装在塑料培养皿或玻璃容器中的培养基。培养基生产商会尽量采用同一生物来源的原料，但这些原料不可避免会有差异，因此需要考虑到培养基批次之间的差异。另外化验室制备的或者生产商制备的培养

34、基的性能很大程度度上取决于培养条件和存贮条件。Impropermediapreparationcancauseunsatisfactoryconditionsformicrobialgrowthorrecoveryandunreliableresults.培养基制备不当会导致微生物生长或回收条件不理想，从而导致不可靠的结果。Therefore,qualitycontroltestsshouldbeperformedonallpreparedmedia,includingmediaassociatedwithswabsormediainstripsandothernontraditionalfo

35、rmats.Testsroutinelyperformedonin-housepreparedmediashouldincludepH,growthpromotion,inhibition,andindicativeproperties(asappropriate),andperiodicstabilitycheckstoconfirmtheexpirationdating.因此，需要对制备的所有培养基进行质量控制检测，包括擦拭结合培养基、条状培养基和其它非传统方式。一般公司自制培养基检测应包括pH值、促生长试验、抑制试验和指示性试验(如需要)，并进行定期稳定性检查以确认有效期。Whenin-

36、housepreparedmicrobiologicalmediaareproperlypreparedandsterilizedusingavalidatedmethod,thegrowth-promotiontestingmaybelimitedtoeachincominglotofdehydratedmedia,unlessotherwiseinstructedbytherelevantcompendialmethod.Ifthemediapreparationprocedurewasnotvalidated,theneverybatchofmediashouldbesubjectedt

37、ogrowth-promotiontesting.Testorganismsmaybeselectedfromtheappropriatecompendialtestchapter.Inaddition,microorganismsusedingrowth-promotiontestingmaybebasedonthemanufacturersrecommendationforaparticularmedium,ormayincluderepresentativeenvironmentalisolates(buttheselatterarenottobeconstruedascompendia

38、lrequirements).如果公司采用经过验证的方法进行培养基制备和灭菌，则促生产试验可以是只针对购入的每个培养基干粉批次，除非相关药典方法另有要求。如果培养基制备程序未经验证，则所制备的每个批次培养基均需要进行促生长试验。检验用菌种可以从相应的药典检测章节中选择。另外，对于特殊的培养基，用于促生长试验的菌株可以采用生产商推荐的品种，或包括有代表性的环境隔离物（但后者不构成药典要求的内容）。Expirationdatesonmediashouldhavesupportinggrowth-promotiontestingtoindicatethattheperformanceofthemed

39、iastillmeetsacceptancecriteriauptoandincludingtheexpirationdate.Thelengthofshelflifeofabatchofmediawilldependonthestabilityoftheingredientsandformulationunderspecifiedconditions,aswellasthetypeofcontainerandclosure.培养基的有效期应能支持促生长试验，在培养基有效期内及到到有效期时，该培养基应仍符合可接受标准。一个批次培养基货架期长度取决于配料成份的稳定性、在特定条件下的配方，以及容器

40、和密封的形式。Whenabatchofmediadoesnotmeettherequirementsofgrowth-promotiontesting,aninvestigationshouldbeinitiatedtoidentifythecause.Thisinvestigationshouldincludeacorrectiveactionplantopreventtherecurrenceoftheproblem.Anybatchofmediathatfailsgrowth-promotiontestingisunsuitableforuse.如果培养基的促生长试验失败，则应启动调查程

41、序寻找原因。该调查应包括纠正措施以防止相同问题重复发生。所有促生产试验失败的培养基不得用于检测。NOTEFailedgrowth-promotiontestresultsmaynotbeusedtonegatepositivetestresults.注：促生产试验失败可能会导致阳性检测为阴性。Somereagentsareusedfordiagnosticpurposestohelpsupportidentificationofmicrobialorganisms,e.g.,Gramstainandoxidasetestreagents.Thesemayhaveattributesthatca

42、nbequalitycontroltestedsimilartomicrobiologicalmedia.Selectthecorrectqualitycontrolstandardmicroorganisms,followingthemanufacturersinstructions,andperformthetestingbeforeunknownsamplediagnostictesting.Allrelevantdiagnosticreagentsshouldbesubjectedtoincomingqualityconfirmationbeforeuse.有些诊断试剂用于帮助鉴别微生

43、物种类，例如革兰氏染色和氧化酶检测试剂。这些试剂具有与微生物培养基类似的进行质量控制测试的特性。选择正确的质量控制标准微生物，按照生产商指示，在未知样品诊断检测前进行测试。所有相关的诊断试剂均应进行进厂质量确认才可使用。Specialcareshouldbetakenwithmediathatisusedinsterilitytests(seeSterilityTests71forrequirements)andinenvironmentalmonitoringstudies.Mediausedforenvironmentalmonitoringofcriticalareasshouldpre

44、ferablybedouble-wrappedandterminallysterilized.Ifterminalsterilizationisnotperformed,mediashouldbesubjectedto100%pre-incubationandinspectionbeforeusewithinacriticalarea.NOTEGrowth-promotiontestingforthismediamustbeperformedafterthepreincubationstage.Thiswillpreventextraneouscontaminationfrombeingcar

45、riedintocontrolledenvironmentsandwillpreventfalse-positiveresults.Araisedagarlevelforsurfacecontactplatesshouldbeverified.用于无菌检测(见71无菌检测)和环境监控研究的培养基需要特别注意。用于关键区域环境监控的培养基最好用双层包装，并采用最终灭菌方式。如果无法进行最终灭菌，则在使用前应进行100%预培养并检查【注：该培养基的促生长试验必须在预培养阶段后进行】。这样能防止由于携入受控制环境中所产生的额外污染，防止假阳性结果。接触碟培养基厚度应进行确认。MAINTENANCEO

46、FMICROBIOLOGICALCULTURES微生物培养维护Biologicalspecimenscanbethemostdelicatestandardstohandlebecausetheirviabilityandcharacteristicsaredependentonadequatehandlingandstorage.Standardizingthehandlingandstorageofculturesbytheuserlaboratoryshouldbedoneinawaythatwillminimizetheopportunityforcontaminationoralte

47、rationofgrowthcharacteristics.Thecarefulandconsistenttreatmentofstockculturesiscriticallyimportanttotheconsistencyofmicrobiologicaltestresults.Culturesforuseincompendialtestsshouldbeacquiredfromanationalculturecollectionoraqualifiedsecondarysupplier.生物种属可能是最精致的分类标准，因为其活性和属性取决于充分的处理和存贮。在使用菌种的化验室，其菌种处

48、理和存贮应采用标准化程序以最大程度降低污染或生长特性变异。对库存菌种进行小心的处理对于保证微生物检验结果的一致性是至关重要的。在药典检测中使用的菌种应从国家菌种保藏中心或有资质的二级供应商处获取。Theycanbeacquiredfrozen,freeze-dried,onslants,orinready-to-useforms.Confirmationofthepurityofthecultureandtheidentityofthecultureshouldbeperformedbeforeitsuseinqualitycontroltesting.菌种可以是冰冻、冻干、斜面或即用即配形式

49、。在将其用于质量控制检测前应对培养物的纯度进行确认和鉴别。Ready-to-useculturesshouldbesubjectedtoincomingtestingforpurityandidentitybeforeuse.Theconfirmationofidentityforcommonlyusedlaboratorystrainsshouldideallybedoneatthelevelofgenusandspecies.临用现配菌种应在进厂时检测其纯度和鉴别方可使用。化验室常用菌株的鉴别最好在其属和种的层次进行。Preparationandresuscitationofculture

50、sshouldfollowtheinstructionsofthesupplieroravalidated,establishedmethod.The“Seed-Lot”techniqueisrecommendedforstorageofstockcultures.菌种的制备和接种应遵守供应商的指示，或遵守经过验证的已有的方法。建议对菌种保藏采用“种子批”技术。Theoriginalsamplefromthenationalculturecollectionoraqualifiedsecondarysupplierisresuscitatedandgrowninanappropriatemed

51、ium.Aliquotsofthisstockculture(thefirsttransferorpassage)aresuspendedinacryoprotectivemedium,transferredtovials,andfrozenat一30Corbelow,untiluse.Ifstoredat-70C,orinlyophilizedform,strainsmaybekeptindefinitely.Thesefrozenstockscanthenbeusedtoinoculatemonthlyorweeklyworkingcultures.Onceopened,donotrefr

52、eezeunusedcellsuspensionsafterculturingaworkingsuspension.Theunusedportionshouldbediscardedtominimizetheriskoflossofviabilityandcontaminationofthestock.从国家菌种保藏中心或有资质的二级供应商获得的原始样品在适当的培养基中进行复苏和生产。库存菌种(第一次传递或通道)的分装为低温防护培养基形成混悬液，转移至小瓶，在-30C下冷冻直至使用。如果在-70C下存贮，或以冻干形式存贮，菌种可以无限期地存贮。这些冷冻存贮的菌种可以用作每周或每月的工作菌种。未

53、使用的部分应该弃去以最大程度降低活性损失的风险和存贮污染的风险。Thenumberoftransfersofworkingcontrolculturesshouldbetrackedtopreventexcessivesubculturingthatincreasestheriskofphenotypicalterationormutation.Thenumberoftransfersallowableforspecificcompendialtestsmaybespecifiedinthattest.Onepassageisdefinedasthetransferoforganismsfro

54、maviableculturetoafreshmediumwithgrowthofthemicroorganisms.Anyformofsubculturingisconsideredtobeatransfer/passage.工作控制菌种的传代应可以追溯，以防止过度传代，从而增加表型变异性或异化的风险。特定的药典检测所允许传代的次数必须在该检测中指明。一种途径可以界定为从活菌传代至新鲜培养基，在其中微生物可以生长。任何形式的接种都应被作为是传代/通道。LABORATORYEQUIPMENT实验室设备Mostequipment(incubators,waterbaths,andautoclav

55、es)issubjecttostandardvalidationpracticesofincomingqualification,operationalqualification,andperformancequalification.Additionally,periodiccalibration(generallyannually)iscommonlyrequired.Newequipment,criticaltotheoperationofthelaboratory,shouldbequalifiedaccordingtoaprotocolapprovedbythequalityassu

56、ranceunit(QAU).Inaddition,regularcleaningandsanitizationofequipmentsuchasincubators,refrigerators,andwaterbathsshouldbeperformedtominimizethepotentialforcontaminationinthelaboratory.Doorsealsofincubatorsandrefrigeratorsshouldbecleanedandcheckedforstateofrepair.主要设备(培养箱、水浴锅、灭菌器)需要进行标准验证，包括进厂确认、操作确认和性

57、能确认。另外，一般还需要对其进行周期性校正(一般每年校正)。化验室操作的关键新设备应根据质量部门批准的方案进行确认。此外，对设备，如培养箱、冰箱和水浴锅，进行定期清洁和灭菌都是需要的，这样可以最大程度降低化验室中污染的风险。培养箱和冰箱的门封应进行清洁并对维修状态进行检查。Instruments(pHmetersandspectrophotometers)usedinamicrobiologylaboratoryshouldbecalibratedonaregularscheduleandtestedtoverifyperformanceonaroutinebasis.Thefrequency

58、ofcalibrationandperformanceverificationwillvarybasedonthetypeofinstrumentandtheimportanceofthatequipmenttothegenerationofdatainthelaboratory.在微生物化验室使用的仪器（pH计和光度计）应按计划进行定期校正，并测试以按照常规要求确认其性能。校正频率和性能确认可以根据仪器的类型和仪器重要性而制订不同的周期。Equipmentthatisdifficulttosanitize(suchasrefrigeratorsandincubators)shouldbede

59、dicatedtoasepticoperations(suchasstorageofmediafortestingandincubationofsterilitytestsamples)andlivecultureoperationstominimizethepotentialforinadvertentcontaminationofthetests.难以灭菌的设备(例如冰箱和培养箱)，如果用于无菌操作(例如测试用培养基存贮和无菌测试样品的培养)和活菌操作，则分开专用，以最大程度降低可能无意的污染。Autoclavesarecentraltotheoperationofthelaborator

60、yandmusthavepropervalidationinplacetodemonstrateadequatesterilizationforavarietyofoperations.Autoclaveresourcesmustbeavailable(andvalidated)tosterilizewastemedia(ifperformedinthatlaboratory)aswellasthemediapreparedinthatlaboratory.Thechoiceofoneorseveralautoclavesisnotdrivenbyaneedtoseparateaseptica