Lawrence Livermore National Laboratory, Livermore, California, United States of America.
Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo, Japan.
PLoS One. 2018 Dec 28;13(12):e0209683. doi: 10.1371/journal.pone.0209683. eCollection 2018.
Kawasaki disease (KD), first identified in 1967, is a pediatric vasculitis of unknown etiology that has an increasing incidence in Japan and many other countries. KD can cause coronary artery aneurysms. Its epidemiological characteristics, such as seasonality and clinical picture of acute systemic inflammation with prodromal intestinal/respiratory symptoms, suggest an infectious etiology for KD. Interestingly, multiple host genotypes have been identified as predisposing factors for KD. To explore experimental methodology for identifying etiological agent(s) for KD and to optimize epidemiological study design (particularly the sample size) for future studies, we conducted a pilot study. For a 1-year period, we prospectively enrolled 11 patients with KD. To each KD patient, we assigned two control individuals (one with diarrhea and the other with respiratory infections), matched for age, sex, and season of diagnosis. During the acute phase of disease, we collected peripheral blood, nasopharyngeal aspirate, and feces. We also determined genotypes, to identify those that confer susceptibility to KD. There was no statistically significant difference in the frequency of the risk genotypes between KD patients and control subjects. We also used unbiased metagenomic sequencing to analyze these samples. Metagenomic sequencing and PCR detected torque teno virus 7 (TTV7) in two patients with KD (18%), but not in control subjects (P = 0.111). Sanger sequencing revealed that the TTV7 found in the two KD patients contained almost identical variants in nucleotide and identical changes in resulting amino acid, relative to the reference sequence. Additionally, we estimated the sample size that would be required to demonstrate a statistical correlation between TTV7 and KD. Future larger scale studies with carefully optimized metagenomic sequencing experiments and adequate sample size are warranted to further examine the association between KD and potential pathogens, including TTV7.
川崎病(KD)于 1967 年首次被发现,是一种病因不明的儿科血管炎,在日本和许多其他国家的发病率呈上升趋势。KD 可导致冠状动脉瘤。其流行病学特征,如季节性和伴有前驱肠道/呼吸道症状的急性全身炎症的临床表现,提示 KD 具有感染病因。有趣的是,已鉴定出多种宿主基因型为 KD 的易患因素。为了探索鉴定 KD 病因剂的实验方法学,并优化未来研究的流行病学研究设计(特别是样本量),我们进行了一项试点研究。在为期 1 年的时间里,我们前瞻性地招募了 11 名 KD 患者。我们为每位 KD 患者分配了两名对照个体(一名患有腹泻,另一名患有呼吸道感染),匹配年龄、性别和诊断季节。在疾病的急性期,我们采集了外周血、鼻咽抽吸物和粪便。我们还确定了基因型,以鉴定那些易患 KD 的基因型。KD 患者和对照个体之间风险基因型的频率没有统计学差异。我们还使用无偏的宏基因组测序来分析这些样本。宏基因组测序和 PCR 在两名 KD 患者(18%)中检测到转矩腾病毒 7(TTV7),但在对照个体中未检测到(P = 0.111)。Sanger 测序显示,在两名 KD 患者中发现的 TTV7 在核苷酸上几乎具有相同的变体,与参考序列相比,在导致的氨基酸上也有相同的变化。此外,我们估计了需要的样本量,以证明 TTV7 与 KD 之间存在统计学相关性。需要进行更大规模的未来研究,优化精心设计的宏基因组测序实验和足够的样本量,以进一步检查 KD 与包括 TTV7 在内的潜在病原体之间的关联。