中国动物检疫
中國動物檢疫
중국동물검역
Chinese Journal of Animal Health Inspection
2015年
9期
77-81
,共5页
赵永刚%邹艳丽%张永强%吴晓东%王清华%王志亮
趙永剛%鄒豔麗%張永彊%吳曉東%王清華%王誌亮
조영강%추염려%장영강%오효동%왕청화%왕지량
苏丹型埃博拉病毒%序列比对%焦磷酸测序
囌丹型埃博拉病毒%序列比對%焦燐痠測序
소단형애박랍병독%서렬비대%초린산측서
Sudan type EBOV virus%sequence comparison%pyrosequencing
[目的]本研究旨在通过对埃博拉病毒进行序列信息分析的基础上,利用焦磷酸测序技术对苏丹型埃博拉病毒进行快速检测和鉴定.[方法]通过序列信息比对,设计苏丹型埃博拉病毒基因保守区段的扩增引物及测序引物.通过人工方法合成一段基因序列,PCR扩增目的基因片段,经体外转录制备cRNA,RT-PCR扩增目的基因片段,采用焦磷酸测序技术针对目的基因进行保守核苷酸区段的测序分析.[结果]通过序列信息比对寻找到苏丹型埃博拉病毒基因型的核苷酸保守区段,经焦磷酸测序后能进一步确证毒株的序列信息为苏丹型埃博拉病毒.经过与华大基因公司进行Sanger法测序比较,结果完全一致.[结论] 基于序列分析的焦磷酸测序技术可以作为进一步确证方法使用.
[目的]本研究旨在通過對埃博拉病毒進行序列信息分析的基礎上,利用焦燐痠測序技術對囌丹型埃博拉病毒進行快速檢測和鑒定.[方法]通過序列信息比對,設計囌丹型埃博拉病毒基因保守區段的擴增引物及測序引物.通過人工方法閤成一段基因序列,PCR擴增目的基因片段,經體外轉錄製備cRNA,RT-PCR擴增目的基因片段,採用焦燐痠測序技術針對目的基因進行保守覈苷痠區段的測序分析.[結果]通過序列信息比對尋找到囌丹型埃博拉病毒基因型的覈苷痠保守區段,經焦燐痠測序後能進一步確證毒株的序列信息為囌丹型埃博拉病毒.經過與華大基因公司進行Sanger法測序比較,結果完全一緻.[結論] 基于序列分析的焦燐痠測序技術可以作為進一步確證方法使用.
[목적]본연구지재통과대애박랍병독진행서렬신식분석적기출상,이용초린산측서기술대소단형애박랍병독진행쾌속검측화감정.[방법]통과서렬신식비대,설계소단형애박랍병독기인보수구단적확증인물급측서인물.통과인공방법합성일단기인서렬,PCR확증목적기인편단,경체외전록제비cRNA,RT-PCR확증목적기인편단,채용초린산측서기술침대목적기인진행보수핵감산구단적측서분석.[결과]통과서렬신식비대심조도소단형애박랍병독기인형적핵감산보수구단,경초린산측서후능진일보학증독주적서렬신식위소단형애박랍병독.경과여화대기인공사진행Sanger법측서비교,결과완전일치.[결론] 기우서렬분석적초린산측서기술가이작위진일보학증방법사용.
[Objective] To establish a new molecular method for simultaneous and rapid detection and confirmation of Ebola virus by using sequence analysis combined with pyrosequencing technology. [Methods] Amplification primers and a sequencing primer were designed according to the published sequences of conserved L gene regions of three Sudan Ebola viruses in GenBank. A gene sequence was synthesized artificially, and the target gene were amplified by PCR, to prepare cRNA through in vitro transcription. RT-PCR was developed for conserved regions of the gene, and the RT-PCR products were pysrosequenced.[Results] The characterized nucleotide segment was obtained by sequence analysis, and then the segment was confirmed as Sudan Ebola virus. Compared to the Sanger method of Huada, they showed 100% agreement.[Conclusion] Pyrosequencing technology based on sequence analysis can be used to confirm Sudan Ebola virus.