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Updated: Jun 15, 2025
Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Unveiling the gut-heart potential connection: microbiota's role in kawasaki disease and coronary artery lesions
Qing Yang1, Wei Tang1, Jiayu Ren1
1Department of Pediatrics, Qilu Hospital of Shandong University, Jinan, China.
Insights
Gut microbiota dysbiosis is linked to coronary artery lesions (CALs) in children with Kawasaki disease (KD). Microbiota-targeted strategies may improve outcomes for KD patients with CALs.
Area of Science:
- Microbiology
- Immunology
- Pediatrics
Background:
- Kawasaki disease (KD) is a leading cause of acquired heart disease in children, characterized by systemic immune vasculitis.
- Emerging research highlights the gut microbiota's role in the gut-vascular axis and its potential influence on coronary artery lesions (CALs) in KD.
- Understanding this connection is crucial for developing novel therapeutic approaches.
Purpose of the Study:
- To investigate the association between gut microbiota composition and the development of CALs in pediatric patients with KD.
- To identify specific bacterial biomarkers indicative of CALs in both acute and non-acute phases of KD.
- To explore the potential of microbiota-targeted interventions for managing CALs.
Main Methods:
- Analysis of gut microbiota composition in 60 children with KD and 30 healthy controls using 16S rRNA sequencing.
- Assessment of alpha diversity indices and application of t-tests to compare microbial communities.
- Utilizing Linear discriminant analysis Effect Size (LEfSe) to identify microbial biomarkers associated with CALs.
Main Results:
- Children with KD in the acute phase exhibited reduced gut microbial diversity and richness, with an overgrowth of opportunistic pathogens.
- Specific bacterial genera, including *Burkholderia-Caballeronia-Paraburkholderia* and *Hungatella*, were enriched in KD patients with CALs.
- In the non-acute phase, CALs were associated with depletion of beneficial bacteria like *Anaerostipes* and *Subdoligranulum*, and enrichment of others such as *Coprobacillus*.
Conclusions:
- Gut microbiota dysbiosis plays a significant role in the pathogenesis of CALs in Kawasaki disease.
- The identified bacterial biomarkers could aid in predicting or diagnosing CALs.
- Microbiota-modulating therapies hold promise for mitigating CALs and improving long-term outcomes in KD patients.
Background:
Kawasaki disease (KD) is an acute systemic immune vasculitis predominantly affecting medium and small arteries, commonly observed in pediatric patients. It represents the most common form of acquired heart disease in this population. Emerging evidence suggests that gut microbiota can modulate the gut-vascular axis, influencing coronary artery lesions (CALs). This study aims to elucidate the potential association between gut microbiota, KD, and CALs, as well as identify bacterial biomarkers for CALs.
Methods:
We analyzed the gut microbiota composition of 60 children with KD (15 in the acute phase, 45 in the nonacute phase) and 30 healthy controls using alpha diversity indices and t-tests. Microbial biomarkers were identified through LEfSe to analyze the interplay between gut microbiota and CALs in KD during acute and non-acute phases.
Results:
In the acute phase, KD children exhibited decreased richness and diversity of gut microbiota, characterized by dysbiosis, particularly a reduction in short-chain fatty acid-producing bacteria and overgrowth of opportunistic pathogens. Thirteen genera showed statistically significant changes, including Enterococcus, Bacteroides, Faecalibacterium, Agathobacter, Lachnospiraceae_NK4A136_group, Roseburia, Monoglobus, etc. LEfSe analysis revealed enrichment of Burkholderia-Caballeronia-Paraburkholderia, Hungatella, and Clostridium_innocuum group in patients with concurrent CALs, while Halomonas was depleted. In the nonacute phase, gut microbiota diversity was similar to healthy controls, but Streptococcus was upregulated, while Eubacterium_eligens_group and Erysipelotrichaceae_UCG_003 were downregulated. CALs were associated with reduced Anaerostipes, Subdoligranulum, Roseburia, and Lachnospira. LEfSe analysis also showed enrichment of Coprobacillus, Paludicola, Lautropia, UCG-009, Acetanaerobacterium, Methylobacterium-Methylorubrum, and Alistipes in patients with CALs.
Conclusions:
Our findings provide evidence of gut microbiota dysbiosis in KD children, suggesting its involvement in CALs development. It indicates that under the premise of standardized treatment during the acute phase of KD, it is plausible that microbiota-targeted strategies may alleviate CALs, thereby improving patient prognosis.
