PRV感染PK-15细胞对prv-miR-LLT11a表达时相的影响

摘要

为研究伪狂犬病病毒(PRV)的致病机制,将PRV QBA株按0.5个感染复数(MOI)的剂量接种PK-15细胞,分别在感染后0、1、2、4、6、8、12、24 h收取细胞并提取microRNA(miRNA),采用茎环引物实时荧光定量PCR(RT-qPCR)检测prv-miR-LLT11a的表达时相.RT-qPCR扩增结果显示,熔解曲线峰值单一,扩增曲线拐点清晰.RT-qPCR检测结果表明,PRV QBA株感染PK-15细胞1 h后,prv-miR-LLT11a表达量极显著升高,随后表达量下调,在感染6 h后表达量降至最低,感染8 h后表达量持续急剧升高.%In order to study the pathogenic mechanism of pseudorabies virus(PRV),PK-15 cells were infected with PRV QBA strain at 0.5 MOI,and cells were harvested and microRNA(miRNA) were extracted at 0,1,2,4,6,8,12,24 hours post infection(hpi).Then the temporal expression of prv-miR-LLT11a was detected by stem-loop real-time quantitative PCR(RT-qPCR).The amplification results of RT-qPCR showed that the melting curve had a single peak and a clear inflection point.RT-qPCR data showed that the expression of prv-miR-LLT11a remarkably significant increased in PK-15 cells after 1 hour infection with PRV QBA strain,then decreased.Its expression level decreased to the lowest level at 6 hpi,and increased significantly after 8 hpi.