title: Identification of Cereulide-Producing Bacillus cereus by Nucleic Acid Chromatography and Reverse Transcription Real-Time PCR.
abstract: RNA extracts were analyzed with a nucleic acid sequence-based amplification (NASBA) - nucleic acid chromatography and a reverse transcription-quantitative PCR assay (RT-qPCR) based on the TaqMan probe for identification of cereulide-producing Bacillus cereus. All 100 emetic B. cereus strains were found to give positive results, but 50 diarrheal B. cereus strains and other bacterial species showed negative results in the NASBA-chromatography. That is, the assay could selectively identify the emetic strains among B. cereus strains. Also, the B. cereus contents of more than 10(7) cfu/ml were required for the identification of the cereulide-producing strains in this assay. In qRT-PCR assays, all 100 emetic type strains of B. cereus produced 10(2) - 10(4) copy numbers per ng of the RNA preparation, and the strains produced 10(4) copies including ones which had the high vacuolation activities of HEp-2 cells.
authors.affiliation: Laboratory of Hygiene, Kagawa Nutrition University.
authors.forename: Shigeko
authors.initials: S
authors.lastname: Ueda
authors.forename: Manami
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Kayoko
authors.initials: K
authors.lastname: Eguchi
authors.forename: Miki
authors.initials: M
authors.lastname: Iwase
mesh_terms: D001409:Bacillus cereus / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D002845:Chromatography; D047630:Depsipeptides / Q000096:biosynthesis*; D012329:RNA, Bacterial / Q000737:chemistry* / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D047630:Depsipeptides; D012329:RNA, Bacterial; C089294:cereulide