title: Development of a high-resolution melting-based approach for efficient differentiation among Bacillus cereus group isolates.
abstract: Strains belonging to Bacillus cereus Group include six different species, among which are Bacillus thuringiensis, Bacillus weihenstephanensis, and Bacillus cereus sensu stricto, a causative agent of food poisoning. Sequence of the panC-housekeeping gene is used for B. cereus Group affiliation to seven major phylogenetic groups (I-VII) with different ecological niches and variations in thermal growth range and spore heat resistance of B. cereus Group microorganisms varies among phylogenetic groups. We assigned a selection of B. cereus sensu stricto strains related to food poisoning from the Spanish cultivar Collection (Valencia) to Group IV strains based on panC gene sequence. Thermal inactivation assays revealed variability of spore heat resistance within these Group IV strains. Adequate food sanitizing treatments therefore require fast and reliable identification of particular strains. In the present study, feasibility of genotyping via high-resolution melting (HRM) analysis was examined. HRM analysis of amplified polymorphic 16S-23 intergenic spacer region (ISR) region proved to be discriminatory for B. cereus sensu stricto strain typing, while two other polymorphic regions within the bacterial rRNA operon allowed differentiation between Bacillus species, demonstrating its applicability for discrimination on the species and strain level within B. cereus Group.
authors.affiliation: Departamento Ingeniería Alimentos y del Equipamiento Agrícola, Escuela Técnica Superior de Ingeniería Agronómica, Universidad Politécnica de Cartagena, Cartagena, Spain.
authors.forename: Vera
authors.initials: V
authors.lastname: Antolinos
authors.forename: Pablo S
authors.initials: PS
authors.lastname: Fernández
authors.forename: María
authors.initials: M
authors.lastname: Ros-Chumillas
authors.forename: Paula M
authors.initials: PM
authors.lastname: Periago
authors.forename: Julia
authors.initials: J
authors.lastname: Weiss
mesh_terms: D001407:Bacillus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001409:Bacillus cereus / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D001413:Bacillus thuringiensis / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D021901:DNA, Intergenic / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005240:Feasibility Studies; D005513:Food Inspection / Q000379:methods; D005516:Food Microbiology; D005517:Foodborne Diseases / Q000517:prevention & control; D006358:Hot Temperature; D050296:Microbial Viability; D058889:Molecular Typing / Q000379:methods*; D009691:Nucleic Acid Denaturation; D010453:Peptide Synthases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012335:RNA, Ribosomal / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013171:Spores, Bacterial / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D021901:DNA, Intergenic; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D010453:Peptide Synthases; C022318:pantothenate synthetase