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1、PCR实验室基本要求1.1.1PCR实验室基本要求2.1PCR实验室依据所使用的方法可分为试剂准备、标本制备、扩增、产物分析等三或四个区域。只是使用实时荧光PCR仪、HIV病毒载量测定仪的PCR实验室�有上述前面三个区域即可。各区域应完全独立分隔�不应有任何的空气直通。BasedonthemethodsusedbyPCRlab,thePCRlaboratorycanbedividedintothreeorfourroomsorareasfor(i)reagentpreparation,(ii)samplepreparation,(iii)amplificati
2、onarea,and(iv)productanalysisarea.IfthePCRlaboratoryonlyusesreal-timefluorescencePCRinstrumentsand/orinstrumentsforHIVvirusload,thePCRlabmayhaveonlythefirstthreeroomsorareas(reagentpreparation,samplepreparation,amplification).TheroomsorareasofthePCRlabshouldbeindependentlyseparated,
3、andthereisnoairflowthroughtheseparatedroomsorareas.2.2产物分析区或三个区域的最后一个区域�即扩增及产物分析区��空气流向应由室外向室内�可通过在室内设置通风橱、排风扇或其他排风系统达到空气流由室外向室内流动的要求。ForPCRproductanalysisareaorlastareaofthreeareas(areaforamplificationandproductanalysis),directionofairflowshallbefromoutsidetoinsideoftheareabysettin
4、gupfumehood,exhaustfanorotherexhaustsysteminthearea.2.3PCR实验室各区域仪器设备配备通常如下�TheequipmentconfigurationforPCRlabsareasfollows:2.3.1试剂准备区:Reagentpreparationarea仪器设备主要应有加样器、冰箱、天平、低速离心机、混匀器、可移动紫外灯等。可使用超净工作台作为试剂配制操作台面。Themainequipmenthereshouldbe:pipette,refrigerator,balance,lowspeedcentri
5、fuge,vortexmixer,mobileUVlamp,etc.ItcanuseLaminarflowcabinetforthereagentpreparation.2.3.2标本制备区samplepreparationarea仪器设备主要应有生物安全柜�最好为B2,可避免提取核酸在柜内反复循环�造成标本间交叉“污染”�出现假阳性结果。此外还应配备加样器、台式高速离心机�冷冻及常温�、台式低速离心机、恒温设备�水浴和/或干浴仪�、冰箱、混匀器和可移动紫外灯等。Themainequipmentisthebio-safetycabinet(typeB2isbe
6、tter,becauseitcanavoidrecyclingofnucleicacidcabinetandcrosscontaminationofspecimens).Besides,itshallbeequippedwithpipettes,centrifugetablehighspeed(refrigeratedandnormaltemperature),centrifugetablelowspeed,constanttemperaturedevices(waterbathordrybath),refrigerator,vortexmixerandmob
7、ilelampUVetc.2.3.3扩增区amplificationarea主要仪器就是核酸扩增热循环仪�PCR仪�实时荧光或普通的�。热循环仪的电源应专用�并配备一个稳压电源或UPS�以防止由于电压的波动对扩增测定的影响。此外�根据工作需要�还可配备加样器、超净台等。Themainequipmenthereisathermalcyclersystemfornucleicacidamplification(PCR,quantitativefluorescencedetectionorcommon).Powerforthethermalcyclershallbe
8、providedwithamanost
