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孔梅, 段卫平, 陈锦英, 李颖, 李晓燕, 苏旭, 王撷秀. 乙型流感病毒的RT-PCR-SSCP法初筛分类[J]. 中国公共卫生, 2006, 22(10): 1229-1230. DOI: 10.11847/zgggws2006-22-10-44
引用本文: 孔梅, 段卫平, 陈锦英, 李颖, 李晓燕, 苏旭, 王撷秀. 乙型流感病毒的RT-PCR-SSCP法初筛分类[J]. 中国公共卫生, 2006, 22(10): 1229-1230. DOI: 10.11847/zgggws2006-22-10-44
KONG Mei, DUAN Weiping, CHEN Jinying, . Typing of influenza B viruses HA1 gene by RT-PCR-SSCP[J]. Chinese Journal of Public Health, 2006, 22(10): 1229-1230. DOI: 10.11847/zgggws2006-22-10-44
Citation: KONG Mei, DUAN Weiping, CHEN Jinying, . Typing of influenza B viruses HA1 gene by RT-PCR-SSCP[J]. Chinese Journal of Public Health, 2006, 22(10): 1229-1230. DOI: 10.11847/zgggws2006-22-10-44

乙型流感病毒的RT-PCR-SSCP法初筛分类

Typing of influenza B viruses HA1 gene by RT-PCR-SSCP

  • 摘要:
      目的   应用单链构象多态性(SSCP)对乙型流感病毒血凝素重链区(HA1)基因抗原决定簇集中区进行突变筛选, 了解2001~2004年天津地区流行株HA1基因的变异特性。
      方法   病毒培养液提取RNA后RT-PCR扩增HA1基因易发生变异308 bp片段, 经SSCP分类, 选取各类代表株PCR产物纯化后进行核苷酸序列测定, 用DNAS-TAR软件分析。
      结果   天津地区近几年分离的乙型流感病毒根据SSCP电泳图谱不同分为5类。Victoria系易变区抗原表位有6个位点氨基酸不同于B/Hongkong/330/2001, Yamagata系易变区抗原表位有3个位点氨基酸不同于B/Beijing/76/98, 有10个位点不同于B/Sichuan/379/99。Yamagata系与Victoria系相比缺失第163位氨基酸天冬酰胺(N)。O相与D相毒株间未发现氨基酸有差异。
      结论   RCR-SSCP方法可作为初步筛查流感病毒代表株的方法。近几年天津地区乙型流感病毒两大谱系交替流行, 且抗原性进一步发生漂移, Yamagata系毒株为B/Beijing/76/98类似株。

     

    Abstract:
      Objective   To screen the influenza B viruses mutation within the amplified region which included the high-mutable site by RT-PCR-SSCP, and to study the characteristics of HA1 gene of influenza B viruses isolated from 2001 to 2004 in Tianjin.
      Methods   Viral RNA was used for RT-PCR to amplify the high-mutable region of 308bp, then was classified by SSCP.The purified PCR products of representative strains were sequenced.The data obtained were analysed with the software DNASTAR.
      Results   Influenza B viruses isolated in Tianjin in recent years were classified into five sorts by SSCP.Victoria-lineage strains compared with B/Hongkong/330/2001 had six mutation points in antigenic deter minants within amplified region, Yamagata-lineage strains compared with B/Beijing/76/98 had three mutation points, and with B/Sichuan/379/99 had ten.Yamagata-lineage strains deleted mutation at amino acid 163(N).There was no difference of amino acid on HA1 gene high-mutable site with O or D phase feature.
      Conclusion   RT-PCR-SSCP can be applied for screening the representative strains of influenza virus.Yamagata-lineage and Victoria-lineage strains circulated alternatively in Tianjin recently, and the substitution of amino acids occurred gradually.Yamag ata-lineage strains were closely related to B/Beijing/76/98-like.

     

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