Association between Clostridioides difficile ribotypes, restriction endonuclease analysis types, and toxin gene expression. 2018

Hiroki Watanabe, and Yusuke Koizumi, and Asami Matsumoto, and Nobuhiro Asai, and Yuka Yamagishi, and Hiroshige Mikamo
Department of Clinical Infectious Diseases, Aichi Medical University Hospital, 1-1, Yazako-karimata, Nagakute, Aichi 480-1195, Japan; Department of Infection Control and Prevention, Aichi Medical University Hospital, 1-1, Yazako-karimata, Nagakute, Aichi 480-1195, Japan.

OBJECTIVE Clostridioides difficile strains cause severe infection. Previous studies suggested that the virulence of C. difficile is dependent on ribotype; however, this hypothesis is still controversial. We aim to investigate the relationship between ribotypes, restriction endonuclease analysis (REA) types, and toxin gene expression in C. difficile strains. METHODS We utilized 53 clinical C. difficile strains. All strains were assigned a molecular strain type using PCR ribotyping and REA typing and classified into 17 ribotypes and six REA types. The expression of toxin genes (tcdA, tcdB, and cdtB) in C. difficile strains were quantified by real-time PCR using each specific primer set, and expression was normalized to that of the housekeeping gene rpoA. RESULTS All 53 strains expressed tcdB and four strains expressed cdtB. Five strains did not express tcdA. Most ribotype and REA type strains expressed tcdA and tcdB similar to the BAA-1870 strain. In cdtB-positive strains, the cdtB expression levels were similar to those in the BAA-1870 strain. tcdA and tcdB expression levels were similar in the cdtB-positive and cdtB-negative strains. CONCLUSIONS Toxin gene expression was not associated with the ribotype. Production of binary toxin C. difficile transferase was not related to tcdA and tcdB expression levels.

UI MeSH Term Description Entries
D012150 Polymorphism, Restriction Fragment Length Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment. RFLP,Restriction Fragment Length Polymorphism,RFLPs,Restriction Fragment Length Polymorphisms
D003015 Clostridium Infections Infections with bacteria of the genus CLOSTRIDIUM and closely related CLOSTRIDIOIDES species. Clostridioides Infections,Clostridioides difficile Infection,Clostridioides perfringens Food Poisoning,Clostridioides perfringens Infections,Clostridioides sordellii Infection,Clostridium difficile Infections,Clostridium sordellii Infections,Clostridium difficile Infection,Clostridium perfringens Food Poisoning,Clostridium perfringens Infections,Clostridium sordellii Infection,Infections, Clostridium,Clostridioides Infection,Clostridioides perfringens Infection,Clostridium Infection,Clostridium perfringens Infection,Infection, Clostridioides difficile,Infection, Clostridioides sordellii,Infection, Clostridium,Infection, Clostridium difficile,Infection, Clostridium sordellii
D004262 DNA Restriction Enzymes Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1. Restriction Endonucleases,DNA Restriction Enzyme,Restriction Endonuclease,Endonuclease, Restriction,Endonucleases, Restriction,Enzymes, DNA Restriction,Restriction Enzyme, DNA,Restriction Enzymes, DNA
D004768 Enterotoxins Substances that are toxic to the intestinal tract causing vomiting, diarrhea, etc.; most common enterotoxins are produced by bacteria. Staphylococcal Enterotoxin,Enterotoxin,Staphylococcal Enterotoxins,Enterotoxin, Staphylococcal,Enterotoxins, Staphylococcal
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000091402 Prohibitins Ubiquitously expressed conserved proteins associated with multiple functions including APOPTOSIS; CELL PROLIFERATION, regulations of various immune and mitochondrial functions, and cellular stress responses. Prohibitin 1 and prohibitin 2 form a ring-shaped complex in the INNER MITOCHONDRIAL MEMBRANE. Prohibitin
D001426 Bacterial Proteins Proteins found in any species of bacterium. Bacterial Gene Products,Bacterial Gene Proteins,Gene Products, Bacterial,Bacterial Gene Product,Bacterial Gene Protein,Bacterial Protein,Gene Product, Bacterial,Gene Protein, Bacterial,Gene Proteins, Bacterial,Protein, Bacterial,Proteins, Bacterial
D001427 Bacterial Toxins Toxic substances formed in or elaborated by bacteria; they are usually proteins with high molecular weight and antigenicity; some are used as antibiotics and some to skin test for the presence of or susceptibility to certain diseases. Bacterial Toxin,Toxins, Bacterial,Toxin, Bacterial
D015183 Restriction Mapping Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA. Endonuclease Mapping, Restriction,Enzyme Mapping, Restriction,Site Mapping, Restriction,Analysis, Restriction Enzyme,Enzyme Analysis, Restriction,Restriction Enzyme Analysis,Analyses, Restriction Enzyme,Endonuclease Mappings, Restriction,Enzyme Analyses, Restriction,Enzyme Mappings, Restriction,Mapping, Restriction,Mapping, Restriction Endonuclease,Mapping, Restriction Enzyme,Mapping, Restriction Site,Mappings, Restriction,Mappings, Restriction Endonuclease,Mappings, Restriction Enzyme,Mappings, Restriction Site,Restriction Endonuclease Mapping,Restriction Endonuclease Mappings,Restriction Enzyme Analyses,Restriction Enzyme Mapping,Restriction Enzyme Mappings,Restriction Mappings,Restriction Site Mapping,Restriction Site Mappings,Site Mappings, Restriction
D016360 Clostridioides difficile A common inhabitant of the colon flora in human infants and sometimes in adults. The type species Clostridioides difficile is formerly known as Clostridium difficile. It is a causative agent for CLOSTRIDIOIDES INFECTIONS and is associated with PSEUDOMEMBRANOUS ENTEROCOLITIS in patients receiving antibiotic therapy. Clostridium difficile

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