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Author Spotlight: Leptospira DNA Detection in Water for Environmental Analysis and Disease Surveillance
Published on: June 14, 2024
Comparative genome characterization of Leptospira interrogans from mild and severe leptospirosis patients
Songtham Anuntakarun1, Vorthon Sawaswong1, Rungrat Jitvaropas2
1Program in Bioinformatics and Computational Biology, Graduate School, Chulalongkorn University, Bangkok 10330, Thailand.
Abstract:
Leptospirosis is a zoonotic disease caused by spirochetes from the genus Leptospira. In Thailand, Leptospira interrogans is a major cause of leptospirosis. Leptospirosis patients present with a wide range of clinical manifestations from asymptomatic, mild infections to severe illness involving organ failure. For better understanding the difference between Leptospira isolates causing mild and severe leptospirosis, illumina sequencing was used to sequence genomic DNA in both serotypes. DNA of Leptospira isolated from two patients, one with mild and another with severe symptoms, were included in this study. The paired-end reads were removed adapters and trimmed with Q30 score using Trimmomatic. Trimmed reads were constructed to contigs and scaffolds using SPAdes. Cross-contamination of scaffolds was evaluated by ContEst16s. Prokka tool for bacterial annotation was used to annotate sequences from both Leptospira isolates. Predicted amino acid sequences from Prokka were searched in EggNOG and David gene ontology database to characterize gene ontology. In addition, Leptospira from mild and severe patients, that passed the criteria e-value < 10e-5 from blastP against virulence factor database, were used to analyze with Venn diagram. From this study, we found 13 and 12 genes that were unique in the isolates from mild and severe patients, respectively. The 12 genes in the severe isolate might be virulence factor genes that affect disease severity. However, these genes should be validated in further study.
Insights
Genomic sequencing of Leptospira isolates revealed unique genes in both mild and severe leptospirosis cases. The severe cases had 12 unique genes, potentially indicating virulence factors that warrant further investigation.
Area of Science:
- Microbiology
- Genomics
- Infectious Diseases
Background:
- Leptospirosis is a zoonotic disease caused by Leptospira spirochetes.
- Leptospira interrogans is a significant cause of leptospirosis in Thailand.
- Clinical presentations range from mild to severe organ failure.
Purpose of the Study:
- To investigate genomic differences between Leptospira isolates from mild and severe leptospirosis patients.
- To identify potential virulence factors associated with disease severity.
Main Methods:
- Whole-genome sequencing using Illumina technology.
- Bioinformatic analyses including read trimming, assembly, annotation (Prokka), and gene ontology characterization (EggNOG, DAVID).
- Virulence factor identification using BLASTP and Venn diagram analysis.
Main Results:
- 13 unique genes were identified in the isolate from a mild case.
- 12 unique genes were identified in the isolate from a severe case.
- The unique genes in the severe isolate are potential virulence factors.
Conclusions:
- Genomic analysis can differentiate Leptospira isolates causing varying disease severities.
- The 12 unique genes in the severe isolate may contribute to leptospirosis severity.
- Further validation of these potential virulence genes is recommended.
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