欢迎访问《中国循证儿科杂志》官方网站，今天是
English Version
中国循证儿科杂志

中国循证儿科杂志 ›› 2024, Vol. 19 ›› Issue (1): 12-18.DOI: 10.3969/j.issn.1673-5501.2024.01.003

• 论著 • 上一篇    下一篇

基于病毒组学的川崎病潜在病毒感染特征分析

林沙1a,1b　刘晓亮1a,1b　华益民1a,1b　范振鑫2　吴刚1a,1b　李一飞1a,1b   

  1. 1 四川大学华西第二医院　成都，610041，a 出生缺陷与相关妇儿疾病教育部重点实验室，b 四川大学华西第二医院儿童心血管科；2 四川大学生命科学学院　成都，610041
  • 收稿日期:2023-11-16 修回日期:2023-12-11 出版日期:2024-02-25 发布日期:2024-02-25
  • 通讯作者: 吴刚，李一飞

Metagenomics analysis of the association between Kawasaki diseases and viral infection

LIN Sha1a,1b, LIU Xiaoliang1a,1b, HUA Yimin1a,1b, FAN Zhenxin2, WU Gang1a,1b, LI Yifei1a,1b   

  1. 1 West China Second University Hospital Sichuan University, Chengdu 610041, China, a Ministry of Education Key Laboratory of Women and Children's Diseases and Birth Defects; b Department of Pediatrics, Ministry of Education Key Laboratory of Women and Children's Diseases and Birth Defects; 2 College of Life Sciences, Sichuan University, Chengdu 610041, China
  • Received:2023-11-16 Revised:2023-12-11 Online:2024-02-25 Published:2024-02-25
  • Contact: WU Gang，LI Yifei

PDF

54

摘要： 背景 目前虽然有许多研究从流行病学、遗传学、感染、免疫、炎症等方面探讨川崎病（KD）病因，但仍无定论。 目的 通过对KD患儿血液的微生物组学和病毒组学的测序分析，尝试从微生物学和病毒学角度分析KD可能的病因，揭示KD患儿体内微生物和病毒的组成及相对丰度，并寻找KD的潜在致病病毒。 设计 病例对照研究。 方法 采集KD和健康儿童血液标本，行病毒样颗粒的纯化、提取和测序，并对宏基因组测序和原始数据质量控制，通过MEGAHIT软件实现宏基因组的组装，运用kraken2进行物种注释，获得样本的物种在各个分类层级（门、目、科、属、种）的相对丰度，利用在线工具Wekemo Bioincloud将分类群丰度按相对丰度标准化，并将前20个分类群进行展示，利用线性判别分析进行比较，以线性回归分析（LDA）>2为标准筛选组间在丰度上有显著差异的物种。 主要结局指标 物种在门、目、科、属、种的相对丰度。 结果 在微生物组学层面，KD组与对照组极为相似，只有科萨病毒门和芽孢杆菌Y1的相对丰度差异有统计学意义。特异性病毒组学高通量测序分析显示，门级：KD组与对照组相比，Uroviricota、核质病毒门和杆状病毒门的相对丰度明显降低；属水平：Orthohepadnavirus、Pegunavirus、Montyvirus、正内罗病毒属、Toursvirus等多个属的相对丰度较对照组显著增加；种水平：KD组Woodchuck hepatitis virus（WHV）、丙酸杆菌病毒（PHL112N00, ATCC29399BT，Pirate和P105）、Diadromus pulchellus toursvirus（DpTV-1）等的相对丰度明显高于对照组。 结论 本研究筛选出了几种候选的KD致病性病毒，如WHV、Orthonairovirus和DpTV-1；推测这些病毒可能通过破坏免疫系统导致KD患儿出现急性发热或肝损伤，但还需要更多的研究证实。

关键词: 川崎病, 病毒感染, 宏基因组学, 高通量测序, 生物信息学

Abstract: Background Despite that numerous researches have investigated the pathogenesis of Kawasaki disease (KD), including epidemiology, genetics, infection, immunity, and inflammation. However, the causes of KD is still not well underlined. Objective We analyzed the blood samples of KD patients and control cases using modified highthroughput metagenomics sequencing to identify the bacterial and viral composition and relative abundances, aiming to demonstrate the potential pathogenic virus of KD. Design Casecontrol study. Methods We collected the blood samples from KD patients and controls, then purification, extraction and sequencing of viruslike particles had been completed. After quality control of raw data, macrogenome was assembled by MEGAHIT software and species annotating was performed on kraken2. Obtaining relative abundance of each sample at different taxonomic levels (phylum, order, family, genus, and species), we normalized taxon abundance with the online tool Wekemo Bioincloud. The top 20 taxa were displayed and analyzed comparatively using linear discriminant analysis. LDA>2 was set as the criteria of significant differences in abundance between groups for filtering potential pathogenic species. Main outcome measures Relative abundance of each sample at different taxonomic levels (phylum, order, family, genus, and species). Results We found that there was little difference between KDs and the controls in terms of overall microorganism. Only phylum Cossaviricota and species Bacillussp Y1 were significantly different. However, there was a great significant difference between viral population annotations. The relative abundances of Uroviricota, Nucleocytoviricota, and Taleaviricota were significantly higher in controls compared with KD patients. Various genera, such as Orthohepadnavirus, Pegunavirus, Montyvirus, Orthonairovirus, Toursvirus, showed significantly higher relative abundance in KDs. In KD patients, the relative abundance of Woodchuckhepatitis virus, Propionibacterium virus (PHL112N00, ATCC29399BT, Pirate, and P105), Diadromus pulchellus toursvirus, and so on were significantly more abundant than that in controls. Conclusion Summarily, we screened out several candidates KD pathogenic viruses, like WHV, Orthonairovirus, and DpTV1. These viruses may be responsible for acute febrile illness or liver damage in KD patients, or they may be able to disrupt patients' immune systems, but further investigations are required.

Key words: Kawasaki disease, Virus infection, Metagenomics, High-throughput sequencing, Bioinformatics