中国兽医科学2013，Vol.43Issue(8)：788-794,7.
猪细小病毒四川株的分离鉴定及其一步生长曲线
Isolation and identification of porcine parvovirus strain SC-L and its one-step growth curve
摘要
Abstract
To study the proliferation characteristics of porcine parvovirus(PPV),a strain of virus was isolated from the liver,kidney and mesenteric lymph node of the abortion piglet using PK-15 cells,and the isolated virus was proved to be PPV,named SC-L,through PCR detection,plaque purification,physical and chemical properties experiment,microneutralization test and hemagglutination test.According to the VP1 and NS1 genes of PPV,a pair of specific primers was designed,and real-time fluorescent quantitative PCR (FQ-PCR) was introduced to quantify PPV.The SC-L isolate was used to infect PK-15 cells,and the infected cells and supernatant were collected at different time points post-infection.FQ-PCR was used to analyze PPV DNA quantity in different samples,and one-step growth curves were then drawn.At 4-20 h post infection,PPV DNA of extracellular virus increased slowly,virus DNA employed a logarithmic growth after 20 h and reached the maximum at 56 h.Afterwards the virus content decreased rapidly and tended to be stable at 68 h.At 4 h post-infection,microdosage PPV DNA of intracellular virus was detected,the virus content of intracellular virus increase slowly at 4-24 h post-infection,virus DNA employed a logarithmic growth after 24 h and reached the maximum at 44 h.Afterwards the virus content decreased slowly.While PPV multiplied to a certain number in PK-15 cells,virion was released from the infected cells and the maximum extracellular virion was higher than that of intracellular one.关键词
猪细小病毒/实时荧光定量PCR/一步生长曲线Key words
porcine parvovirus/real-time fluorescent quantitative PCR/one-step growth curve分类
农业科技引用本文复制引用
江一帆,吴云飞,朱玲,徐志文,阳爱国,廖珊,陈蕾,黄仆,郭万柱..猪细小病毒四川株的分离鉴定及其一步生长曲线[J].中国兽医科学,2013,43(8):788-794,7.基金项目
四川省青年基金项目(09ZQ026-030) （09ZQ026-030）
教育部新世纪优秀人才支撑计划项目(NCET 11-1059) （NCET 11-1059）
四川省科技支撑项目(2010NZ0112,2012NZ0001) （2010NZ0112,2012NZ0001）
