日本臨床微生物学雑誌第22巻第3号
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- よしじろう いちぬの
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1 2012 STEC STEC 1) 1) 1) 2) 2) 3) 1) 2) 3) (Enterohemorrhagic Escherichia coli; EHEC) STEC XM-EHEC 5S A /ViEHEC O157, O26, O111 3 STEC, , XM-EHEC, , ViEHEC, EHEC STEC, 2.7, XM-EHEC, 19.8, ViEHEC, 25.6 STEC STEC (O103 O121 O145) STEC EHEC Key words: STEC (Enterohemorrhagic Escherichia coli; EHEC) VT, stx EHEC 1) 4) O157 : H7 O26 : H11, O111 : H- O103, O121, O145 5) Tel: Fax: nagaokanishi-lab275@sutokukai.or.jp EHEC O157 EHEC EHEC STEC XM-EHEC 5S A /ViEHEC EHEC STEC STEC (Lot No. 0T0921) XM-EHEC (Lot No. 008) 5S A /ViEHEC (Lot No.09004) EHEC EHEC 5S A /ViEHEC 5S A SS Vol. 22 No
2 232 ViEHEC 316 Table 1 EHEC 225 O157; 86 O26; 38 O111; 18 O103; 7 O121; 8 O145; 5 EHEC No EHEC O157; ATCC EHEC 9 O157 3 O26 3 O McFarland No μl EHEC O157; ATCC EHEC 9 O157 3 O26 3 O Miles & Misra 6 McFarland No μl 900 μl μl Blue STEC, XM-EHEC, ViEHEC Miles & Misra 4. 1 (EHEC O157; ATCC35150) 3 EHEC 3 O157, O26, O CFU/g Blue CFU/ml 0.5 ml 0.5 g 10 μl STEC, XM-EHEC, ViEHEC 10 μl EHEC EHEC / EHEC Ehly EHEC IKL LIM VP 7 Neg Combo 6.11J SIEMENS 6. STEC Table STEC 10 μl EHEC UV E-hly EHEC STEC EHEC EHEC / Loopamp O 1. Fig. 1 STEC EHEC O157, EHEC O26, EHEC O111 EHEC O157 UV 26 Vol. 22 No
3 STEC 233 Table 1. List of tested strains Strains Serover Origin VT N Subtotal EHEC Escherichia coli O157 : H7 O157 : H ATCC VT1, 2 1 VT1, 2 14 Isolates VT2 60 VT1 9 Isolates VT1, 2 1 VT2 1 O26 : H11 Isolates VT1, 2 1 VT1 30 VT1, 2 2 O26 : H Isolates VT1 5 O111 : H11 Isolate VT1 1 VT1, 2 4 O111 : H Isolates VT1 13 O103 : H Isolates VT1 2 O103 : H2 Isolates VT1 4 O103 : H11 Isolate VT1 1 O121 : H19 Isolates VT2 7 O121 : H14 Isolate VT2 1 O145 : H Isolates VT2 3 VT2 1 O145 : H NT Isolates VT1 1 Others* Isolates VT1, 2 3 VT2 18 VT1 42 ATCC O6 : H12 Isolates 2 O18 : H7 Isolate 1 O26 : H Isolate 1 O124 : H- Isolate 1 O157 : H Isolate 1 Klebsiella oxytoca ATCC Klebsiella pneumoniae NCTC Enterobacter cloacae ATCC Citrobacter freundii ATCC Proteus mirabilis ATCC Pseudomonas aeruginosa ATCC Bacillus subtilis ATCC Staphylococcus aureus ATCC Enterococcus faecalis ATCC Candida albicans ATCC Other bacteria** Isolates 74 Total 316 NT, not tested; OUT, 0-untypeable * O1 (2), O8 (1), O22 (1), O25 (1), O41 (1), O63 (3), O76 (2), O91 (26), O113 (2), O115 (4), O118 (1), O119 (1), O128 (1), O130 (1), O143 (1), O148 (1), O150 (1), O165 (7), OUT (5) ** Escherichia fergusonii (3), C. freundii (6), E. cloacae (53), Edwardsiella tarda (1), K. pneumoniae (1), K. oxytoca (1), Klebsiella ornithinolytica (1), P. mirabilis (1), Providencia rettegeri (1), P. aeruginosa (2), B. subtilis (1), S. aureus (1), E. faecalis (1), C. albicans (1) Vol. 22 No
4 234 Fig. 1. Representative observation on CHROMagar STEC, XM-EHEC and ViEHEC EHEC O157 : H7 ATCC 35150, EHEC O26 : H11 isolate and EHEC O111 : H- isolate were incubated at 35 for 24 hours. 28 Vol. 22 No
5 STEC 235 Table 2. Recovery rate of CHROMagar STEC, XM-EHEC and ViEHEC with Modified DRIGALSKI agar Blue Tested strains Recovery rate ( ) CHROM XM-EHEC ViEHEC EHEC O157 : H7 ATCC Isolate Isolate Isolate EHEC O26 : H11 Isolate Isolate Isolate EHEC O111 : H Isolate Isolate Isolate Mean CHROM, CHROMagar STEC XM-EHEC EHEC O157 EHEC O26 EHEC O111 ViEHEC O157 EHEC O26 EHEC O Table 2 STEC, XM-EHEC, ViEHEC EHEC O , , , EHEC O , , , EHEC O , , STEC Enterobacter cloacae CFU/ml O157 O26 4. Table 3 STEC XM-EHEC CFU/g ViEHEC 10 2 CFU/g EHEC STEC EHEC 9 2.7, XM-EHEC , ViEHEC E-hly Table 4 XM-EHEC ViEHEC O , 17.0 E. coli, Enterobacter Klebsiella 6. STEC (O157, O26, O111) EHEC STEC EHEC (O103, O121, O145) EHEC 63.5 ( Table 5) 7. EHEC EHEC 16 (0.007 ; 16/228,829) EHEC O157 1 ( ) EHEC O26 8 ( ) 7 ( ) EHEC O157 : H7 O26 : H11, O111 : H 3 3 XM-EHEC ViEH- EC E. coli Enterobacter Vol. 22 No
6 236 Table 3. Detection of EHEC in fecal samples inoculated with EHEC O157, O26, or O111 Inoculated strains CFU/g (EHEC in feces) Detection from DRIGALSKI CHROM XM-EHEC ViEHEC EHEC O157 : H7 EHEC O26 : H11 EHEC O111 : H ATCC Isolate Isolate Isolate Pos Pos (16) Pos (16) Neg Pos Pos Pos Pos Pos Pos Pos Pos Pos Neg Neg Neg Pos Pos (15) Pos (23) Pos (5) Pos Pos Pos Pos Pos Pos (10) Pos (11) Neg Pos Pos Pos Pos Pos Pos Pos Pos Pos Pos (20) Pos (5) Neg Pos Pos Pos Pos Pos Pos Pos Pos CHROM, CHROMagar STEC; DRIGALSKI, Modified DRIGALSKI agar Blue; Pos, Positive; Neg, Negative The numbers in parenthesis are the numbers of colonies from evaluated culture media. Table 4. Detection rate and enterohemolysin production of EHEC-like colonies in fecal samples N 324 Colony color of CHROMagar STEC XM-EHEC ViEHEC Mauve MUG ( ) Mauve, MUG ( ) Violet Blue Pale pink/ Pink Clear/ Brown Green/ Blue Pale pink/ Violet Escherichia coli Enterobacter cloacae E. agglomerans Klebsiella pneumoniae K. oxytoca Citrobacter spp Proteus mirabilis 1 1 P. vulgaris 1 7 Pseudomonas aeruginosa 2 6 Pseudomonas spp. 4 Acinetobacter spp. 1 Staphylococcus aureus 1 Subtotal ( ) 4 (1.2) 5 (1.5) 29 (9.0) 1 (0.3) 34 (10.5) 25 (7.7) 3 (0.9) 55 (17.0) Total ( ) 9 (2.7) 64 (19.8) 83 (25.6) Enterohemolysis ( ) 0 (0.0) 0 (0.0) 0 (0.0) MUG, 4-methylumbelliferyl-β-d-glucuronide;, positive;, negative Klebsiella 19.8, 25.6 STEC EHEC EHEC O157, O26, O103, O111, O121, O145 5) STEC (Table 5) 30 Vol. 22 No
7 STEC 237 Strain Table 5. Colony observation of EHEC on CHROMagar STEC and enterohemolysin agar Mauve MUG( ) Colony observation Enterohemolysis Mauve, No growth Blue Clear Pos Neg MUG( ) EHEC O EHEC O EHEC O EHEC O EHEC O EHEC O Other EHEC Other Escherichia coli Other bacteria 3* 1** MUG, 4-methylumbelliferyl-β-d-glucuronide;, positive; -, negative; Pos, Positive; Neg, Negative * Enterobacter cloacae (2), Citrobacter freundii (1) ** Escherichia hermanii (1) 2010 EHEC O157 1,078 EHEC O EHEC , 19.1, 13.2EHEC O157 1 ( ) EHEC O26 8 ( ) EHEC 7 ( ) EHEC O157 CT-SMAC EHEC O157 EHEC 8) 9) STEC 9 EHEC enterohemolysis EHEC 63.5 EHEC eaea 10) Fukushima EHEC eaea EHEC CT- SMAC 11) (O157, O26, O111, O103, O121, O145) Tzschoppe STEC terb 12) STEC CT-SMAC (data not shown) 2010 (stx) O104 O ,400 STEC Tzschoppe 12 stx VT1/VT2 ELISA Loopamp EHEC EHEC 13) STEC EHEC EHEC Vol. 22 No
8 238 STEC 1) Edelman, R., M. A. Karmali, P. A. Fleming From the National Institutes of Health. Summary of the International Symposium and Workshop on Infections due to Verocytotoxin (Shiga-like toxin)-producing Escherichia coli. J. Infect. Dis. 157: ) Karmali, M. A., M. Petric, C. Lim, et al The association between idiopathic hemolytic uremic syndrome and infection by verotoxin-producing Escherichia coli. J. Infect. Dis. 151: ) Pai, C. H., N. Ahmed, H. Lior, et al Epidemiology of sporadic diarrhea due to verocytotoxin-producing Escherichia coli: A two-year prospective study. J. Infect. Dis. 157: ) Riley, L. W., R. S. Remis, S. D. Helgerson, et al Hemorrhagic colitis associated with a rare Escherichia coli serotype. N. Engl. J. Med. 308: ) Gyles, C. L Shiga toxin-producing Escherichia coli: An overview. J. Anim. Sci. 85: E45 E62. 6) 1978 p , 7) p ) Beutin, L., M. Montenegro, I. Φrskov, et al Close association of verotoxin (shiga-like toxin) production with enterohemolysin production in strains of Escherichia coli. J. Clin. Microbiol. 27: ) : ) 2010 p ) Fukusima, H., K. Hiroshina, M. Gomyoda Selective isolation of eae-positive strains of shigatoxinproducing Eschericha coli. J. Clin. Microbiol. 38: ) Tzschoppe, M., A. Martin, L. Beutin A rapid procedure for the detection and isolation of enterohaemorrhagic Escherichia coli (EHEC) serogroup O26, O103, O111, O118, O121, O145 and O157 strains and the aggregative EHEC O104: H4 strain from ready-to-eat vegetables. Int. J. Food. Microbiol. 152: ) 2010 O26 O157 (SEL) 20: Comparison of Three Chromogenic Culture Medium for Differentiation of Enterohemorrhagic Escherichia coli (EHEC), and Availability of CHROMagar STEC for EHEC Carrier-Screening Yuuki Sumiya, 1) Masanori Umezawa, 1) Hiroshi Yamazaki, 1) Mitsuru Okamoto, 2) Mizuho Hayashida, 2) Takamasa Kaneko 3) 1) Nagaoka-Nishi Hospital 2) Tokyo Kenbikyoin Foundation 3) Kanto Chemical Co., Inc. Chromogenic medium CHROMagar STEC and XM-EHEC and ViEHEC were evaluated as selective isolation media for enterohemorrhagic Escherichia coli (EHEC). The growth support abilities (percentages) of three sera groups mainly in Japan (O157, O26, O111) were in CHROMagar STEC, in XM-EHEC, and in ViEHEC. The EHEC-similar colony appearance rates in fecal samples were 2.7 in CHROMagar STEC, 19.8 in XM- EHEC, and 25.6 in ViEHEC. These findings indicated that CHROMagar STEC showed the highest specificity for EHEC. The detection sensitivity of CHROMagar STEC for the main three sera groups was 100. Moreover, outbreaks have been reported, and mauve colonies were formed in all tested strains with O157, O26, O111, O103, O121, and O145, which are believed to be highly associated with disease. These results indicated that CHROMagar STEC had sufficiently high sensitivity and specificity that it would be suitable medium on the theme of carrier-screening for EHEC. 32 Vol. 22 No
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