GB 19438-3

ICS11.220B41GB/T19438.3—2004H7RT-PCRMethodofreal-timeRT-PCRforthedetectionoftheAvianInfluenzavirussubtypeH72004-2-152004-2-15GB/T19438.3—2004IGB/T1.1—20001GB/T19438.3—20041H7RT-PCR1RT-PCRH7H72GB/T19438.1-2004RT-PCR3RT-PCR-CtCyclethresholdRNARibonucleicacidTaqTaqDNAPBSDEPC4TaqManH75’FAMR3’TAMRAQ5’RQTaq5’→3’RQPCRPCR55.1RNAH7RT-PCR1A75%RNAGB6682-920.01M(pH7.2)PBSGB/T19438.4-2004A121215min1GB/T19438.3—2004210000U/mL5.212000gPCR28-200.5µL10µL5µL20µL20µL200µL200µL1000µL66.1Eppendorf121215min6.26.2.135l.0mLPBSEppendorf6.2.22.0g10mLPBS1000g10minEppendorf6.2.3Eppendorf6.2.42824h-70(3)77.1GB/T19438.1-2004C7.27.2.17.2.27.2.2.1n1.5mLEppendorfGB/T19438.3—200437.2.2.2600µL200µL200µL5s412000g15min7.2.2.37.2.2.11.5mLEppendorf500µL(20)7.2.2.47.2.2.2500µL7.2.2.5412000g15min7.2.2.6600µL75412000g15min7.2.2.74000glOs3minRNA7.2.2.811µLDEPCRNA2000g5sRNA2hRT-PCR-707.37.3.17.3.1.17.3.1.2RTPCRTaq2000g5sPCRn15µLRT-PCR0.25µLTaq0.25nRT-PCRPCR15µL7.3.27.3.2.17.3.2.2PCR7.2.2.8RNAlOµL500g30s7.3.3RT—PCR7.3.3.17.3.3.27.3.2.2PCRPCR7.3.4RT-PCRH742/30min92/3min92/10s45/30s72/1min592/10s60/30s4088.18.1.18.1.28.2Ct28.08.3GB/T19438.3—200448.3.1Ct30H78.3.2CtH78.4Ct30.0CtGB/T—5AH7RT-PCRA.148/30mL1DEPC1mL1H7RT-PCR750µL1RT-PCR1/12Taq12µL11mL1RNA1mL1A.2A.2.1RNA4-20A.2.2DEPC1%DEPCRNAA.2.3RT-PCRA.3RT-PCR