食品製造環境由来の Bacillus cereus の菌株
Contents
1. MLST cereus PET ST ST ST 40 143 156 166 1 22. DNA ExoSAP IT GE Healthcare dNTP 25 fmol DNA DTCS Quick Start Kit Beckman Coulter 30 95 60 70 200 2 CEQ8000 Beckman Coulter 6 MLST MLST START2 The Bacillus cereus Multi Locus Sequence Typing website http pubmlst org bcereus 1 cezezs 14
3. TIFT 114006 ST 40 TIFT 114009 ST 156 7 4 3 1 MLST PCR Priest p ceres MLST gmk 2 14 gz 1 1 2 1 PCR 3 ceezs MLST PCR strains gmk smk F3 glpf ilyD Primer set IivD 2 iv4F pta pur pycA tpi TIFT 114001 TIFT 114002 TIFT 114003 TIFT 114004 TI
4. 29 79 83 2013 79 Bacillus cereus Strain Typing of Bacillus cereus lsolates from Food Plants Atsuhito Enda and Tomoe Enda Bacillus cereus Well known food borne pathogen is also possible causal bacteria of aseptically packaged foods and drinks It is required to asses contaminations by strain typing methods for prevention of spoilage in aseptic food plants Multilocus sequence typing MLST was adopted for phylogenetic analyss of 14 strains of B ce7ez y 1solates Nine novel sequence types STs were found and three pairs of two strains isolated from the same plants were revealed to be very close individually One strain with cereulide production gene was found to belong to the ST frequently isolated from rice The MLST method is evaluated to be useful for control of B cereus in aseptic food plants Key words Bacillus cereus MLST Food plant cereulide Bacillus cereus Clostridium botulinum C pe ge77y
5. 7 cereus 1 cereus 1 1 Bacillus cereus Strain Origin CRS gene cereus PFGE Pulsed Field Gel Electrophoresis RAPD Random Amplified Polymorphic DNA DNA AFLP Amplified Fragment Length P
6. CRS 1 TIFT 114011 13 2 PCR 7 Primers Forward Reverse end glpF GCGTTT GTG CTG GTG TAAGT CTG CAATCG GAAGGAAGAAG 59 gmk ATTTAAGTG AGG AAG GGT AGG GCAATGTTCACCAACCACAA 56 gmk F3 GAGAAGTAG AAG AGG ATT GCT CATC same as gmk 55 iD CGG GGC AAA CAT TAA GAGAA GGT TCT GGT CGT TTC CAT TC 58 IvD 2 AGATCG TAT TAC TGC TAC GG GTT ACC ATT TGT GCATAA CGC 58 ilv4F GCAGAGATT AAAGAT AAGGA same as ilvD 2 50 pta GCAGAG CGT TTA GCAAAAGAA TGC AAT GCG AGT TGC TTC TA 56 pur CTG CTG CGAAAAATC ACAAA CTC ACG ATT CGC TGC AAT AA 56 pycA GCG TTA GGT GGAAAC GAAAG CGC GTC CAAGTT TAT GGA AT 57 tpi GCC CAG TAG CAC TTA GCGAC CCGAAACCG TCAAGAATG AT 58 29 2013 81 2 MLST MLST gmk inD PCR
7. B cereus g 7 gmk ilvD pta pur pye4 i 7 ST Sequence Type ST ST PCR MLST 4 4 gmk 2 1 pur 2 7 1 6 ST TIFT 114008 TIFT 114010 72 42 69 42 88 41 30 TTFT 114013 TIFT 114014 14 6 23 9 4 7 15 2 ST
8. 1 cereus MLST 2 MLST cereus MLST j cereus MLST MLST 1 2 7 3 MLST PFGE RAPD AFLP MLST 1 i A I 3 4 PFGE
9. 3 gz gmk gmk F3 ivD ilvYD_2 iv4F ilvD TIFT 114007 114011 ilvyD_2 ilv4F TIFT 114008 114010 gp TIFT 114008 114009 2 59C 56C 3 MLST MLST MLST allele
10. DNA Bacillus cereus 57 69 72 2007 Hill K K Ticknor L 0 Okinaka R T Asay M Blair H Bliss K A Laker M Pardin gton PE Richardson A P Tonks M Beecher D J Kemp J D Kolste A B Wong A C L Keim P and jackson P J Fluorescent amplified fragment 4 5 length polymorphism analysis of Bacillus anthracis Bacillus cereus and Bacillus thuringiensis 1SOlates Appl Envioron Microbiol 70 2 1068 1080 2004 6 MLST 52 7 209 216 2006 Priest F G Barker M Baillie L W Holmes E C and Maiden M C Population structure and 7 evolution of the Bacillus cereus group 7 Bacteriol 186 23 7959 7970 2004 Jolley K A Feil E J Chan M S and Maiden M C Sequence type analysis and recombinational tests START Bioinformatics 17 1230 1231 2001 8
11. ST ST 26 33 ST 26 TIFT 114011 29 2013 83 33 1 12 35 11 761 777 2007 PET 33 4 161 166 2005 Liu P Y Ke S C and Chen S L Use of pulsed field gel electrophoresis to investigate a Dseudo 2 3 outbreak of Bacillus cereus in a pediatric unit Clin Microbiol 35 6 1533 1535 1997
12. PCR Detection Kit TaKaRa 20 ul 0 1 DNA TAE Tris Acetate EDTA 3 NuSieve 3 1 Agarose Lonza 100V 30 15 15 365 nm 4 MLST PCR MLST Priest DNA glpf gmk ilvD pta 7 pyc4 7 7 2 30 15 02mM 1C 5 PCR
13. FT 114005 TIFT 114006 TIFT 114007 TIFT 114008 TIFT 114009 TIFT 114010 TIFT 114011 TIFT 114012 TIFT 114013 TIFT 114014 5 C 82 29 2013 4 pj cezezxs MLST Allelic profile ST glpf gmk ilvD pia pur pye4 7 TIFT 114001 15 58 10 8 4 7 14 TIFT 114002 46 9 39 29 29 25 7 166 TIFT 114003 15 20 23 8 64 39 TIFT 114004 33 8 40 19 60 17 7 143 TIFT 114005 47 9 14 26 2 36 7 TIFT 114006 39 9 14 12 24 14 7 40 TIFT 114007 19 2 31 17 19 3 2 TIFT 114008 72 42 69 42 88 41 30 TIFT 114009 40 9 14 12 12 25 156 TIFT 114010 72 42 69 42 88 41 30 TIFT 114011 3 2 31 5 16 3 4 26 TIFT 114012 33 8 47 19 8 17 17 TIFT 114013 14 3 9 7 15 TIFT 114014 14 3 9 7 15 5 B cereus MLST i
14. olymorphism MLST Multilocus Sequence Typing cereus TIFT 114001 Aseptic dinkplant TIFT 114002 TIFT 114003 TTFT 114004 a TIFT 114005 TIFT 114006 Aseptic drink plant TIFT 114007 TIFT 114008 a TIFT 114009 TIFT 114010 TIFT 114011 TIFT 114012 2 Aseptic coocked rice plant TIFT 114013 TIFT 114014 a 80 29 2013 2 DNA 35 C p cereus PBS 1 ml DNeasy Blood and Tissue Kit Qiagen DNA 3 DNA Bacillus cereus CRS gene
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