GB 19438-2

ICS11.220B41GB/T19438.2—2004H5RT-PCRMethodofthereal-timeRT-PCRforthedetectionofAvianInfluenzaVirusSubtypeH52004-2-152004-2-15GB/T19438.2—2004IGB/T1.1—20001GB/T19438.2—2004H5RT-PCR1H5RT-PCRH52GB/T19438.1-2004RT-PCR3RT-PCR—CtRNATaqTaqDNAPBSDEPC4TaqManH55’FAMR3’TAMRAQ5’RQTaq5’→3’RQPCRPCRPCR55.1RNA-2075%DEPCGB6682-200.01mol/LpH7.2PBSGB/T19438.4-2004A121215min10000U/mLGB/T19438.2—2004H5RT-PCR1A5.25.2.112000g5.2.2PCR5.2.324-205.2.40.5µL10µL5µL20µL20µL200µL200µL1000µL5.2.55.2.65.2.766.1121215min1602hzzzzEppendorf1.5mL6.26.2.1z35zz1.0mLPBSEppendorf6.2.22.0g10mLPBS10mLPBSEppendorf3000r/min10minEppendorf6.2.3Eppendorf6.32824h-70377.1GB/T19438.1-2004C7.21)GB/T19438.2—20047.2.1n1.5mLEppendorfn7.2.2600µL200µL200µL5s412000r/min15min7.2.37.2.11.5mLEppendorf400L-207.2.2500L7.2.412000r/min15minEppendorf600µL75%7.2.5412000r/min10minEppendorf7.2.64000r/min10sEppendorf3minRNA7.2.711µLDEPCRNA2000r/min5sRNA2hRT-PCR-707.3AIVH5RT-PCRTaq2000r/min5sPCRnnRT-PCRTaq15µL0.25µL0.25nRT-PCRPCR15µL7.4PCR7.2.7RNA10L500r/min30s7.5RT-PCR7.4PCRPCRRT-PCRH542/30min92/3min92/10s45/30s72/1min592/10s60/30s4088.1GB/T19438.2—20048.28.2.1Ct8.2.2Ct28.08.38.3.1Ct8.3.2Ct30.08.3.3Ct30.0CtGB/T19438.2—20041AH5RT-PCRA.148tests/30mL1H5RT-PCR750L1RT-PCRPCR1/12Taq5U/L12L1DEPC1mL11mL1RNA1mL1A.2A.2.1RNA4A.2.2DEPC1%DEPCRNAA.2.3RT-PCRA.3A.3.1A.3.2A.3.3RT-PCRRT-PCRA.3.4-206