快速获取55型腺病毒基因组序列的方法

发布时间：2019-01-05 14:51

【摘要】：【目的】建立快速获取55型腺病毒的全长基因组序列的方法。【方法】根据55型腺病毒的基因组特点,设计覆盖55型腺病毒基因组序列的12对引物,分别以55型腺病毒DNA为模板,扩增得到12个PCR产物,通过对12个PCR产物测序及序列拼接,获得55型腺病毒的全长基因组序列。【结果】从本院急性上呼吸道感染者咽拭子标本中分离得到一株55型腺病毒毒株SF04/SC/2016,以其DNA为模板扩增成功获得12个PCR产物,对其进行测序,并对12段序列进行拼接得到55型腺病毒的全长基因组序列,与已报到的各型腺病毒序列进行比对,采用邻位相连法构建系统发育进化树,所得序列与55型腺病毒处于同一分支,进一步确认该病原体为55型腺病毒。【结论】研究公布的序列和方法,能够实现更方便对腺病毒的快速测序,为揭示55型腺病毒的进化特点及制订疾病防控策略提供了有效手段。
[Abstract]:[objective] to establish a rapid method for obtaining the full length genomic sequence of adenovirus type 55. [methods] according to the genomic characteristics of adenovirus type 55, 12 pairs of primers covering the genome sequence of adenovirus type 55 were designed. Using adenovirus type 55 DNA as template, 12 PCR products were amplified, and 12 PCR products were sequenced and sequenced. The full-length genomic sequence of adenovirus 55 was obtained. [results] 12 PCR products were obtained from throat swabs of patients with acute upper respiratory tract infection by amplification of adenovirus 55 strain SF04/SC/2016, using its DNA as template. The full-length genome sequence of adenovirus type 55 was obtained by sequencing and splicing 12 sequences. The phylogenetic tree of adenovirus type 55 was constructed by means of neighbor linkage method. The obtained sequence is in the same branch as adenovirus type 55, and the pathogen is confirmed as adenovirus type 55. [conclusion] the published sequences and methods can facilitate the rapid sequencing of adenovirus. It provides an effective means to reveal the evolutionary characteristics of adenovirus 55 and to formulate disease prevention and control strategies.
【作者单位】：成都军区总医院检验科;成都军区疾病预防控制中心;
【基金】：国家自然科学基金青年项目(No.81301445) 全军后勤“十二五”重大专项(No.AWS11L009) 全军医学科技青年培育项目(No.14QNP057) 上海市医学生物防护重点实验室开放课题(No.SKLM1401)~~
【分类号】：R373

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