应用不依赖序列的单引物扩增技术获得人博卡病毒全基因序列
응용불의뢰서렬적단인물확증기술획득인박잡병독전기인서렬
Sequence the complete sequence of bocavirus I with SISPA-PCR
  • 万方数据
    中华实验和临床病毒学杂志 중화실험화림상병독학잡지
    CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
    2010年 1期 14-16 ,共3页

    李金松%程卫霞%尧栋萍%兰蓓%虞结梅%刘艳%李永清%章青%靳淼%徐子乾%李丹地%段招军

    리금송%정위하%요동평%란배%우결매%류염%리영청%장청%근묘%서자건%리단지%단초군

    聚合酶链反应%细小病毒科%序列分析 聚閤酶鏈反應%細小病毒科%序列分析
    취합매련반응%세소병독과%서렬분석
    Polymerase chain reaction%Parvoviridae%Sequence analysis
    目的 研究通过不依赖序列的单引物扩增(Sequence independent single primeramplification,SISPA)技术获得腹泻标本中人博卡病毒的全基因序列.方法 筛检仅含有人博卡病毒的标本,经过滤、超速离心富集和核酸酶处理后提取病毒核酸,采用SISPA-PCR技术扩增病毒序列,克隆后测序,经比对分析获得人博卡病毒基因序列,再用DNAstar拼接成全基因序列.结果 获得了人博卡病毒4834bp的序列,仅两端少部分序列未得到.结论 SISPA-PCR技术是一种可直接从样品中获得病毒全基因序列的方法.
    목적 연구통과불의뢰서렬적단인물확증(Sequence independent single primeramplification,SISPA)기술획득복사표본중인박잡병독적전기인서렬.방법 사검부함유인박잡병독적표본,경과려、초속리심부집화핵산매처리후제취병독핵산,채용SISPA-PCR기술확증병독서렬,극륭후측서,경비대분석획득인박잡병독기인서렬,재용DNAstar병접성전기인서렬.결과 획득료인박잡병독4834bp적서렬,부량단소부분서렬미득도.결론 SISPA-PCR기술시일충가직접종양품중획득병독전기인서렬적방법.
    Objective To sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR). Methods To exclude the co-effection samples, all clinical samples of diarrhea cases were screened with special primers of rotavirus, astrovirus, adenovirus, calicivirus and bocavirus I . The virus were enriched through ultracentrifugation. Other nucleic acids,such as human and bacteria genomes, were degradated by DNase I and RNase. DNA of hocavirus was Amplificated with SISPA-PCR,then purificated ,cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar. Results A 4834bp sequence of hocavirus I were assembled. Conclusion SISPA-PCR is an economical and Efficient technique for sequence a virus complete genome.
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