腺病毒55型感染肺炎患者的病原学鉴定及全基因序列分析

发布时间：2018-06-24 08:07

本文选题：不明原因肺炎 + 腺病毒B组型　；参考：《中国病原生物学杂志》2017年03期

【摘要】：目的对1例不明原因肺炎患者进行病原学诊断,分析其病原学特征及重组特点,为其防治提供参考。方法采集患者的咽拭子标本,提取DNA,采用Real-time荧光定量PCR法检测病原体特异性片段;采用Hep-2细胞培养分离病毒;通过Sanger测序hexon基因进行型别鉴定,通过高通量测序技术对病毒进行全基因组测序,采用MEGA6.0软件进行序列比对,通过邻近归并法(Neighbor-Joining)构建种系发生进化树分析基因特征,运用Simplot软件进行序列重组分析。结果从患者咽拭子标本中检测并分离到腺病毒,hexon基因测序后进行比对分析,其与腺病毒B组55型及11型同源性较高;全基因测序显示该病毒与HAdV-55QS-DLL及Shanxi/QZ01/2011株同源性较高,在E1B、DNA-Polymerase、100Ka Hexon和Fiber编码区存在氨基酸变异;种系发生进化树显示该毒株与腺病毒14型同属一支,与腺病毒11型进化分支较近;相似性曲线及BootScan分析显示该病毒株于hexon编码区存在HAdV-11与HAdV-14型间重组。结论本例患者不明原因肺炎由腺病毒B组55型感染引起,该病毒由HAdV-11与HAdV-14重组产生,其生物学特性有待进一步研究。
[Abstract]:Objective to analyze the etiological characteristics and recombination characteristics of a patient with unknown pneumonia and to provide reference for its prevention and treatment. Methods the throat swabs of the patients were collected, DNA was extracted, real-time fluorescence quantitative PCR was used to detect the pathogen specific fragments, Hep-2 cells were cultured to isolate the virus, and the hexon gene was sequenced by Sanger. The whole genome of the virus was sequenced by high-throughput sequencing technique. The sequence alignment was carried out with MEGA 6.0 software. The phylogenetic tree was constructed by neighbor join method and the sequence recombination analysis was carried out with Simplot software. Results the adenovirus hexon gene was sequenced from the throat swabs of the patients, and compared with the adenovirus B group 55 and 11, the whole gene sequencing showed that the virus had high homology with HAdV-55QS-DLL and Shanxi-QZ01 / 2011 strain, and had higher homology with the adenovirus group B group 55 and 11, and the whole gene sequencing showed that the virus had high homology with HAdV-55QS-DLL and Shanxi-QZ01 / 2011 strain. There were amino acid variations in the coding region of 100Ka Hexon and fiber in E1BHN DNA-Polymerase, and the phylogenetic tree showed that the strain belonged to the same branch of adenovirus type 14, and was close to the evolutionary branch of adenovirus type 11. The similarity curve and BootScan analysis showed that there was a recombination between HAdV-11 and HAdV-14 in the hexon coding region. Conclusion the unknown pneumonia of this patient was caused by the infection of adenovirus group B 55, which was produced by the recombination of HAdV-11 and HAdV-14, and its biological characteristics need to be further studied.
【作者单位】：卫生部肠道病原微生物重点实验室江苏省疾病预防控制中心;
【基金】：国家自然科学基金项目(No.81501785) 江苏省自然科学基金项目(No.BK20141030) 江苏省重大新发传染病综合防控科技示范工程项目(No.BE2015714)
【分类号】：R563.1

【相似文献】