利用二代测序技术在不明原因发热的血液样本中检测到人细环病毒  

Detection of human Torque teno virus in serum sample of patients with fever of unknown origin bynext generation sequencing

作　　者：张益[1] 芜为[1] 王佶[1] 申辛欣[1] 何小周[1] 杨梦婕[1] 马学军[1] Zhang Yi;Wu Wei;Wang Ji;Shen Xinxin;He Xiaozhou;Yang Mengfie;Ma Xuejun(Department of Core Facility, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beifing 102206, China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所中心实验室,北京102206

出　　处：《中华实验和临床病毒学杂志》2018年第2期145-149,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：国家重大研究计划（2016TFC1202700,2016YFC1200903）;国家传染病重大专项（2017ZX10302301-004,2017ZX100101,2017ZX10104001）

摘　　要：目的阐明不明原因发热（fever of unknown origin，FUO）的血液样本中用常规方法难以检测到的潜在病原。方法利用二代测序技术序列非依赖的特性对病原核酸进行随机捕获，随后采用多重链置换扩增（multiple displacement amplification, MDA）方法进行富集。在得到测序结果后，进行宏基因组数据分析，构建进化树，最终实现对未知病原的鉴定。结果对10份不明原因发热的血液样本进行宏基因组二代测序分析时未发现引起发热的常见病原体序列，但发现了大量人细环病毒（Torque teno Virus，TTV）存在的现象。将所有相关测序读长进行组装拼接，并计算得到的一致性序列在匹配的参考序列上的基因组覆盖度。在对组装出的置信序列进行进化分析后，发现其分属于α TTV、β TTV和γ TTV三个属。结论对本研究中发现的TTV的基因组特征、进化特征以及作为免疫水平评估的意义进行了分析。而对于本研究中只检测出三个属的TTV的现象，可能是因为TTV具有潜在的病原性以及感染性疾病所导致的宿主免疫水平低下，也有可能是由于其他非感染性病因导致的宿主免疫水平降低所导致。Objective To clarify the potential pathogen for fever of unknown origin （FUO） in serum samples for which pathogenic agents were hardly identified with conventional exainatins. Methods Random capturing the nucleic acid of pathogen was performed by utilizing the property of sequence non- dependence of next generation sequencing （NGS） , followed by enrichment of nucleic acid with multiple displacement amplification （MDA）. After sequencing, metagenomic analysis was applied to the raw data and the phylogenetic tree was built to identify the potential pathogen. Results The result did not indicate common pathogens for FUO but showed the existence of Torque teno Viurs （TTV）. Assembly was carried out to all sequencing reads. The coverage of consensus sequence on reference was calculated. Phylogenetic result indicated that all confidence sequences belonged to 3 genera （ α TTV, β TTV and -γ TTV）. Conclusions The characteristics of genome, phylogenesis of TTV and TTV as signal at immunology level were analyzed and clarified. The possible explanation for detection of TTV in 3 genera may be that TTV itself or non- infectious factors caused the immunosuppression, which finally result e in rise of TTV detection.

关 键 词：不明原因发热 二代测序 宏基因组 人细环病毒 

分 类 号：R440[医药卫生—诊断学] R511[医药卫生—临床医学]