Intestinal microbiota has important effect on severity of hand foot and mouth disease in children

Figures

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  Fig. 1. Enterovirus type identification in HFMDs individual by NGS. A Individuals distribution of the samples with enterovirus type. Red box highlight the co-infection samples. Asterisk means statistical significance difference between mild and severe cases number for enterovirus (Fisher’s exact test p < 0.05). EV71: Human Enterovirus 71; start with “A”: Coxsackievirus A. start with “B”: Coxsackievirus B; start with “EC”: ECHO-virus. B Main virus amount by NGS compare between severe and mild cases. RPM: reads number per million sequenced reads. NS not significant

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  Fig. 2. Enterovirus phylogenetic tree between mild and severe case (VP1). A The phylogenetic tree of the enterovirus 71. B The phylogenetic tree of the coxsackievirus A4. Red color highlighted in branch means severe cases and black means mild case. The branch name started with “H3” or “GZ” was the sequences obtained in this study. The phylogenetic tree was constructed by software MEGA 6 with Maximum Likelihood method

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  Fig. 3. Sample saturation of all sample for gut meta microbe genes. A cumulative sequence data for each sample. B Accumulated samples for all the genes. Each sample number was randomly repeated for 100 times

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  Fig. 4. Severe and mild case index model of Enterovirus positive samples. All 20 genes at lowest error rate: 20.83%. A For each individual, a index was calculated to evaluate the risk of severe HFMD disease (Training: M:S = 104:64). The histogram shows the distribution of indices for all individuals. Red means Mild case and blue means severe case. B Receiver operating characteristic curve of the index model. The area under the ROC curve (AUC) of classifier is 0.9. C new samples for test using the index model (Verify: M:S = 10:10). Mild case and severe case were distinguished by the index model (p < 0.01)

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  Fig. 5. The species enrichment in H, M and S groups of age less than 24 months. White arrow means the group compare. Different circle means different MLG species. The size of the circle indicates abundance of the MLG species. The color of the circle indicates their taxonomic assignment. Connecting lines represent Spearman correlation coefficient values above 0.6 (grey) and below − 0.6 (blue)

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  Fig. 6. MLG species enrichment in 4 groups. A The cluster between species and groups. Each species was compared between groups, one enriched was marker 1, and the most enriched was 3, the range is from 0 to 3. The highlighted text in red represents the microbes which enriched in the server group, and the box section represents the clusters which these microbes belong to. B The pie chart of the genus of MLG species. C The 5 bacterias which different in “M < S > A” in all samples (wilcoxon rank-sum test,*: p < 0.05). Group marker: A: after treatment; H: health; M: mild; S: severe

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  Fig. 7. Heatmap of KEGG function difference between groups. Each function was compared between groups, one enriched was marker 1, and the most enriched was 3, the range is from 0 to 3. Cluster C1 and Cluster C2 were inclined Group H and A Cluster C3 and Cluster C4 were inclined Group M and S. Such as bacterial secretion system and Pathogenicity were enriched in HFMD case