快速获取55型腺病毒基因组序列的方法

Rapid acquisition of complete viral genome sequence of human adenovirus type 55

【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。

[Objective] To build a simple method for rapidly obtaining the full-length genome sequence of adenovirus type 55（HAd V-55）. [Methods] 12 pairs of primers were designed which covered the whole genome sequence of HAd V-55. HAd V-55 DNA was used as the template for PCR and twelve PCR products were obtained, which were sequenced respectively. Resulting sequences were assembled together and then the whole gene sequence was obtained. [Results] A strain of HAd V-55（SF04/SC/2016） was isolated from throat swab specimens of a patient with acute upper respiratory tract infection. Twelve PCR products and their sequences were obtained. The sequences were assembled which yield the full-length genomic sequence. Sequence analysis was performed between this HAd V-55 strain and previous reported HAd V-55 virus in China. Phylogenetic tree was constructed by neighbor-joining method and this virus is in the same branch with previous reported HAd V-55 strains, which confirmed that this pathogen is HAd V-55. [Conclusion] The primer sequences and methods published in this study could facilitate the rapid sequencing of adenovirus, which will be contribute to reveal the evolutionary characteristics of HAd V-55 and the prevention and control of adenovirus type 55 related disease.