学术
猪流感病毒H3N2亚型的分离鉴定与全基因组序列测定
孙志勇[1] 郭万柱[1] 韩国全[1] 陈进会[2] 王小玉[1] 徐志文[1]
[1]四川农业大学动物生物技术中心,雅安625014 [2]东莞出入境检验检疫局综合技术中心,东莞523072
摘 要:
通过鸡胚接种、MDCK细胞培养、血凝及血凝抑制试验、RT-PCR等方法，笔者在四川首次分离并鉴定了1株既能在鸡胚上稳定传代又能在MDCK细胞上稳定产生CPE的H3N2亚型猪流感病毒，命名为A／swine／Sichuan／01／2006（H3N2）；NS基因的测序结果表明：该病毒分离株与A／swine／HongKong／4361／99（H3N2）、A／NewYork／429／2003（H3N2）、A／Queensland／6／2000（H3N2）、A／New South Wales／4／1999（H3N2）等具有代表性的标准毒株的同源性都达到99％；从MDCK细胞中抽提流感病毒基因组进行全基因克隆，构建了11个包含全基因8个基因片段的文库。全基因测序结果表明，A／swine／Siehuan／01／2006（H3N2）共8个节段，全基因序列共计13577bp；基于HA、NA推导蛋白的无根进化树结果显示，四川分离株与人流感标准株A／Queensland／6／2000（H3N2）、A／South Australia／81／2000（H3N2）有较近的亲缘关系，而与猪流感标准株A／swine／Spain／42386／2002（H3N2）的亲缘关系较远。[著者文摘]
文章出处:
《畜牧兽医学报》-2008年39卷2期 -195-200页
栏目信息:
分 类 号:
文献标识码:
A
文章编号:
0366-6964（2008）02-0195-06
Isolation and Identification of Swine Influenza A Subtype H3N2 Strain and Sequencing of the Virus Genome
SUN Zhi-yong , GUO Wan-zhu , HAN Guo-quan , CHEN Jin-hui ,WANG Xiao-yu ,XU Zhi-wen （1. Animal Biotechnology Centre of Sichuan Agricultural University, Yaan 625014,China; 2. Dongguan Entry-Exit Inspection and Quarantine Bureau, Dongguan 523072, China）
Abstract:
A influenza virus strain was isolated from a pig farm in Sichuan province and identified by HA, HI and RT-PCR and some other methods, then it was named A/swine/Sichuan/01/2006 （H3N2）. The virus could not only passage stably in SPF embryonated eggs but also multiply in MDCK cell line and induce conspicuous pathological changes. The sequence analysis indicated that the nucleotide sequence of A/swine/Sichuan/01/2006（H3N2） strain shared 99% homology with standard strains A/swine/Hong Kong/4361/99 （H3N2）, A/New York/429/2003 （H3N2）, A/Queensland/6/ 2000 （H3N2） and A/New South Wales/4/1999 （H3N2）. The results of sequence analysis indicated that the genome of A/swine/Sichuan/01/2006 （H3N2）strain includes 8 fragments and the total sequence is 13 577 bp. Phylogenetic trees based on HA and NA protein gene deduced amino acid sequence showed that A/swine/Sichuan/01/2006 （H3N2 ）have close relationship with the standard strains A/Queensland/6/2000（H3N2） and A/South Australia/ 81 / 2000 （H3N2）.[著者文摘]
Key words:
swine; influenza virus; isolation; identification; genome
基金资助:
“教育部长江学者和创新团队发展计划”项目（IRT0555）
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