In-Depth Analysis of Bacillus anthracis 16S rRNA Genes and Transcripts Reveals Intra- and Intergenomic Diversity and Facilitates Anthrax Detection

• DOI: 10.1128/msystems.01361-21
• Publication Date: 2022-01-25T14:36:06Z
• Authors: Peter Braun; Fee Zimmermann; Mathias C. Walter; Sonja Mantel; Karin Aistleitner; Inga Stürz; Gregor Grass; Kilian Stoecker
• Keywords: Bacillus anthracis

Analysis of 16S rRNA (rRNA) genes provides a central means taxonomic classification bacterial species. Based on presumed sequence identity among species the Bacillus cereus sensu lato group, B. anthracis have been considered unsuitable for diagnosis anthrax pathogen. With recent identification single nucleotide polymorphism in some gene copies, specific becomes feasible. Here, we designed and evaluated set situ, vitro, silico assays to assess unknown state from different perspectives. Using combination digital PCR, fluorescence situ hybridization, long-read genome sequencing, bioinformatics, were able detect quantify unique allele (16S-BA-allele). This was found all available genomes may facilitate differentiation pathogen any close relative. Bioinformatics analysis 959 SRA data sets inferred that abundances genomic arrangements 16S-BA-allele entire operon copy numbers differ considerably between strains. Expression ratios 16S-BA-alleles proportional respective numbers. The findings experimental tools presented here provide detailed insights into intra- intergenomic diversity pave way improved other pathogens with diverse operons. IMPORTANCE For severe infectious diseases, precise detection is crucial antibiotic therapy patient survival. Identification anthracis, causative agent zoonosis anthrax, can be challenging when querying sequences such as small subunit (16S rRNA) genes, which are commonly used typing bacteria. study analyzed broad scale cryptic hitherto underappreciated allelic variant bacterium's their transcripts using significant heterogeneity distribution overall variation uniquely specific, enabled sensitive both DNA transcript levels. methodology likely also applicable otherwise difficult discriminate less harmful relatives.