快速獲取55型腺病毒基因組序列的方法
發(fā)布時間:2019-01-05 14:51
【摘要】:【目的】建立快速獲取55型腺病毒的全長基因組序列的方法。【方法】根據(jù)55型腺病毒的基因組特點,設(shè)計覆蓋55型腺病毒基因組序列的12對引物,分別以55型腺病毒DNA為模板,擴增得到12個PCR產(chǎn)物,通過對12個PCR產(chǎn)物測序及序列拼接,獲得55型腺病毒的全長基因組序列!窘Y(jié)果】從本院急性上呼吸道感染者咽拭子標本中分離得到一株55型腺病毒毒株SF04/SC/2016,以其DNA為模板擴增成功獲得12個PCR產(chǎn)物,對其進行測序,并對12段序列進行拼接得到55型腺病毒的全長基因組序列,與已報到的各型腺病毒序列進行比對,采用鄰位相連法構(gòu)建系統(tǒng)發(fā)育進化樹,所得序列與55型腺病毒處于同一分支,進一步確認該病原體為55型腺病毒!窘Y(jié)論】研究公布的序列和方法,能夠?qū)崿F(xiàn)更方便對腺病毒的快速測序,為揭示55型腺病毒的進化特點及制訂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.
【作者單位】: 成都軍區(qū)總醫(yī)院檢驗科;成都軍區(qū)疾病預(yù)防控制中心;
【基金】:國家自然科學基金青年項目(No.81301445) 全軍后勤“十二五”重大專項(No.AWS11L009) 全軍醫(yī)學科技青年培育項目(No.14QNP057) 上海市醫(yī)學生物防護重點實驗室開放課題(No.SKLM1401)~~
【分類號】:R373
本文編號:2401911
[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.
【作者單位】: 成都軍區(qū)總醫(yī)院檢驗科;成都軍區(qū)疾病預(yù)防控制中心;
【基金】:國家自然科學基金青年項目(No.81301445) 全軍后勤“十二五”重大專項(No.AWS11L009) 全軍醫(yī)學科技青年培育項目(No.14QNP057) 上海市醫(yī)學生物防護重點實驗室開放課題(No.SKLM1401)~~
【分類號】:R373
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