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广州地区猫泛白细胞减少症病毒分离鉴定及全基因组遗传分析

洪马林, 付橙, 黄三, 周沛, 粟硕, 李守军

洪马林, 付橙, 黄三, 周沛, 粟硕, 李守军. 广州地区猫泛白细胞减少症病毒分离鉴定及全基因组遗传分析[J]. 华南农业大学学报, 2017, 38(1): 9-14. DOI: 10.7671/j.issn.1001-411X.2017.01.003
引用本文: 洪马林, 付橙, 黄三, 周沛, 粟硕, 李守军. 广州地区猫泛白细胞减少症病毒分离鉴定及全基因组遗传分析[J]. 华南农业大学学报, 2017, 38(1): 9-14. DOI: 10.7671/j.issn.1001-411X.2017.01.003
HONG Malin, FU Cheng, HUANG San, ZHOU Pei, SU Shuo, LI Shoujun. Isolation, identification and sequence analysis of complete genome of feline panleukopenia virus in Guangzhou area[J]. Journal of South China Agricultural University, 2017, 38(1): 9-14. DOI: 10.7671/j.issn.1001-411X.2017.01.003
Citation: HONG Malin, FU Cheng, HUANG San, ZHOU Pei, SU Shuo, LI Shoujun. Isolation, identification and sequence analysis of complete genome of feline panleukopenia virus in Guangzhou area[J]. Journal of South China Agricultural University, 2017, 38(1): 9-14. DOI: 10.7671/j.issn.1001-411X.2017.01.003

广州地区猫泛白细胞减少症病毒分离鉴定及全基因组遗传分析

基金项目: 

国家重点研发计划 2016YFD0501004

公益性行业(农业)科研专项 201303042

详细信息
    作者简介:

    洪马林(1989—),男,硕士研究生,E-mail:malinhongmark@oxmail.com

    通讯作者:

    李守军(1968—),男,教授,博士,E-mail:shoujunli@scau.edu.cn

  • 中图分类号: S852.65

Isolation, identification and sequence analysis of complete genome of feline panleukopenia virus in Guangzhou area

  • 摘要:
    目的 

    分析广州地区猫泛白细胞减少症病毒(Feline panleukopenia virus, FPLV)的自然重组、跨宿主传播以及流行变异情况。

    方法 

    采集疑似感染猫泛白细胞减少症病毒的猫粪便样品进行细胞分离,并对阳性病料提取基因组DNA,PCR扩增获得病毒的全基因组序列,并与GenBank中相关的参考毒株序列进行遗传进化分析,同时对VP2基因与NS1基因的主要氨基酸位点进行差异分析。

    结果 

    成功获得2株猫泛白细胞减少症病毒及其全基因组序列。遗传进化分析显示,广州地区分离的猫泛白细胞减少症病毒FPLV-GZ01、FPLV-GZ02与我国其他分离毒株FPLV-XJ01、FPLV-HRB属同一分支,提示FPLV-GZ01、FPLV-GZ02由FPLV-XJ1进化而来;NS1基因系统进化树显示,FPLV存在与CPV-447重组的可能。主要氨基酸位点分析显示,FPLV在VP2基因中的主要氨基酸位点上的遗传变异比CPV保守;在NS1基因上发现FPLV-GZ01、FPLV-GZ02分别存在不同程度的氨基酸位点突变。

    结论 

    广州地区分离的猫泛白细胞减少症病毒仍在不断地重组进化。

    Abstract:
    Objective 

    To study the natural recombination, cross host transmission and prevalence of feline panleukopenia virus(FPLV)in Guangzhou area.

    Method 

    Cells were isolated from stool samples of cats seemingly infected with FPLV, genome DNAs were extracted from positive samples, and complete virus genome sequences were amplified by PCR and were compared with relevant reference sequences from GenBank using genetic evolution analysis. The major amino acid sites were analyzed for differences between VP2 gene and NS1 gene.

    Result 

    Two FPLV strains were successfully isolated and the complete genome sequences of FPLV were obtained. The genetic evolution analysis indicated that FPLV-GZ01 and FPLV-GZ02 which were isolated from Guangzhou area belonged to the same branch with the other isolated strains FPLV-XJ1 and FPLV-HRB, and implied FPLV-GZ01 and FPLV-GZ02 evolved from FPLV-XJ01. The NS1 gene phylogenetic analysis demonstrated that FPLV might have recombined with CPV-447. Analysis of major amino acid sites indicated that FPLV was more conservative than CPV in the genetic variation of the VP2 gene. FPLV-GZ01 and FPLV-GZ02 had different degrees of mutation in the amino acid sites of the NS1 gene.

    Conclusion 

    The feline panleukopenia viruses isolated from Guangzhou area have been still recombining and evolving.

  • 图  1   病毒分离结果

    A:对照组的F81细胞;B:试验组的F81细胞。

    Figure  1.   Virus isolation result

    图  2   细小病毒VP2基因的核苷酸序列系统进化树

    Figure  2.   The phylogenetic tree based on nucleotide sequences of parvovirus VP2 gene

    图  3   细小病毒NS1基因的核苷酸序列系统进化树

    Figure  3.   The phylogenetic tree based on nucleotide sequences of parvovirus NS1 gene

    图  4   细小病毒全基因组的核苷酸序列系统进化树

    Figure  4.   The phylogenetic tree based on nucleotide sequences of the complete genomes of parvovirus

    表  1   猫泛白细胞减少症病毒全基因分段引物

    Table  1   Segmented primers for the complete genome of FPLV

    引物名称 引物序列(5′→3′)
    FPLV-1 F:GAATGATAGGCGGTTTGTGTG
    R:TCTTCTGCAATTTCTCTGAGC
    FPLV-2 F:GCAGAAGATAGTGAATGGGTG
    R:TTGGTTGTGTATGTTCAGGTC
    FPLV-3 F:TTATGACAACTAATGAAAATA
    R:TTTTAGTTGGTTTTGTTGGTC
    FPLV-4 F:TGATACACCAGATCATCCATC
    R:TATTTGTTTGCCATGTATGTG
    FPLV-5 F:GAAAATTCTGTGCCAGTACAC
    R:TACCTTTCCACCAAAAATCTG
    FPLV-6 F:AAGACTTCATGTAAATGCACC
    R:AGATTGATACTTATGGTAAGG
    下载: 导出CSV

    表  2   26株细小病毒参考序列

    Table  2   Reference sequences of 26 parvovirus strains

    毒株 登录号 基因型 年份 来源国
    CPV-N M19296 CPV-2 1995 USA
    CPV-M38 M38245 CPV-2 1996 USA
    CPV-F261 FJ005261 CPV-2b 2009 Germany
    CPV-339 AY742933 new CPV-2a 2005 New Zealand
    CPV-447 AY742934 new CPV-2b 2005 USA
    CPV-U6 AY742935 new CPV-2a 2005 Germany
    CPV-395 AY742936 new CPV-2b 2005 USA
    CPV-EF EF011664 new CPV-2a 2006 China
    CPV-NJ01 EU310373 new CPV-2a 2008 China
    CPV-410 EU659119 new CPV-2b 2008 USA
    CPV-13 EU659118 new CPV-2a 2008 USA
    CPV-579 EU659116 CPV-2 2008 USA
    CPV-Y1A D26079 CPV-2a 2008 Japan
    CPV-LZ2 JQ268284 new CPV-2b 2013 China
    CPV-UY349 KM457129 CPV-2c 2014 Uruguay
    CPV-UY135 KM457112 CPV-2c 2014 Uruguay
    FPLV-M382 M38246 FPLV 1996 USA
    FPLV-X55 X55115 FPLV 2005 USA
    FPLV-XJ1 EF988660 FPLV 2007 China
    FPLV-8b EU659114 FPLV 2008 USA
    FPLV-367 EU659111 FPLV 2008 USA
    FPLV-889 EU659113 FPLV 2008 USA
    FPLV-KA EU659115 FPLV 2008 USA
    FPLV-464 EU659112 FPLV 2008 USA
    FPLV-HRB KP280068 FPLV 2015 China
    FPLV-MG132 KP769859 FPLV 2015 Belgium
    下载: 导出CSV

    表  3   VP2基因中主要的氨基酸位点分析

    Table  3   Analysis of major amino acid sites of the VP2 gene

    毒株 氨基酸位点 核酸类型
    80 87 93 103 300 305 323 426 564 568
    FPLV-GZ01 K M K V A D D N N A FPLV
    FPLV-GZ02 K M K V A D D N N A FPLV
    FPLV-M382 K M K V A D D N N A FPLV
    CPV-N R M N A A D N N S G CPV-2
    CPV-Y1A R L N A G Y N N S G CPV-2a
    CPV-NJ01 R L N A G Y N N S G new CPV-2a
    CPV-F261 R L N A G Y N D S G CPV-2b
    CPV-410 R L N A G Y N D S G new CPV-2b
    CPV-UY349 R L N A G Y N E S G CPV-2c
    下载: 导出CSV

    表  4   NS1基因中的部分氨基酸位点分析

    Table  4   Analysis of partial amino acid sites of the NS1 gene

    毒株 氨基酸位点 核酸类型
    248 546 584
    FPLV-GZ01 T P A FPLV
    FPLV-GZ02 T P T FPLV
    FPLV-M382 T S T FPLV
    CPV-N I S T CPV-2
    CPV-Y1A I S T CPV-2a
    CPV-NJ01 I S T new CPV-2a
    CPV-410 I S T new CPV-2b
    CPV-UY349 I S T CPV-2c
    下载: 导出CSV
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出版历程
  • 收稿日期:  2016-03-08
  • 网络出版日期:  2023-05-17
  • 刊出日期:  2017-01-09

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