ῌῦῴ Q̰῎`QQ 2011 25 ῌ῎ ῐ῍ ῧ Q̮Q̲̲QQῥ QQ̯Q b-῾̱̲ῐῴQ῎Ί̯QῥῗΊῬ ῗQῙQQQQῢ CTX-M-27 ῦ̮Q ΅῭῞Ώ1)῎̯Q̱Ὺ1, 2)῎ῖQ̰Q1, 3) 1) ῷQQΊQΊQ̳̳QQQQQ῎̳ QQQQ 2) QQQ̮QQQ̱ ῐ῎̳῎QQ̰ 3) ῷQQΊQ̳Q῭̱QQQ῎QQQQQQQ ῒῌ 22 Ῡ 7 Q 30 ῦQῬ῍ ῌ 23 Ῡ 1 Q 14 ῦQ̲ΐ 2007 Ῡ 12 QῘῪ 2008 Ῡ 2 Qῥ ῟QQ 7 Qῒῥῗ̮ῠ῍ ῧ Q̮QQQῘῪ῾῝῭̯ Escherichia coli 312 QῥῘ῞῍ Clinical and Laboratory Standards Institute QQ̱ῥQ῟̯ ´QQQQQQῗ̰Ῡ double-disk synergy test ΅Q̯̯QQ῍ 15 Q (4.8̮) Ῑ extendedspectrum b-lactamase (ESBL) Q῎Qῡ̮̯̯ῌ ̯ῦ̮̯῍ 13 Qῧ levofloxacin (LVFX) ῥῘ ῞῍ 7 Qῧ LVFX ῗ̰Ῡ trimethoprim/sulfamethoxazole (TMP/SMX) ῥῘ῞ῠ̰ῙQ΅Q ῤ῞ῠῗΎ῍ ESBL Q῎QῦῗQῙQQQQῙQῨ῝῭̯ῌ ῭Ὺ ESBL Q῎ 15 Qῦ ῡQQ ῳῧ῍ 8 QῙ blaCTX-M-27, 4 QῙ blaCTX-M-14 ῡ̮Ύ῍ blaCTX-M-27 ̱̲Qῦ̰̮Ῑ ceftazidime ῥῘ ῞̯QQQ΅Q̯QQῥ̮̯̯ῌ ̰̯῍ ESBL Q῎Qῡ LVFX ῥῘ῞ῙQ΅Q̯ 13 Qῦ̮̯ 7 QῙQ῍Q O25 : H4 ῡ̮Ύ῍ ῼ ῳΌ ῙQῢQ῍QῢῦQ QῙQQ῝῭̯ῌ blaCTX-M ΅̱̲῞̯´῾̱̲Ὸῧῼ 3.4105 cells/recipient ῦQῡῑQῡ῞῞῍ ̯ῦQ TMP/SMX ῙQ̰ΰQῥῡ̰῞ῠ̮̯ῌ ῴQQῥ̰Ύ῍ ῧ Q̮Q̲̲ E. coli ῡῧ῍ CTX-M27 ΅Q῎̯ῬQ῍Q O25 : H4 Ῑ̮ῖ̯̯̮ῬῢῙῸῪῘῢῤ̯̯ῌ ῭ῪῗQῙQ ESBL Q῎QῦῶQ΅Q̮Q̰Ῥ̯̰ῥῧ῍ ῙQQῦQ̳QῠῤQ῏ῗ̰ῩῐQῠῤ̳Q̯QῘῪ῍ ̯ῦ̰Q΅῟ῬῢῙQ̲ῡ̮ῬῢQῖῬῌ Key words: extended-spectrum b-lactamase, uropathogenic Escherichia coli, CTX-M-27, ῗQῙQQ ῏ ῑ ῭Ῥ̰̮ῥῤ̯̯1, 2)ῌ ῤῘῡ̰῍ CTX-M-15 ΅Q῎ Extended-spectrum b-lactamase (ESBL) ῧ῍ class ῞῍ multilocus sequence typing (MLST) 131, Q῍ A b-῾̱̲ῐῴῥ̮῞῍ 3 QῚ̱ῼ` ̱̲ Q O25 : H4 ῢ̮̮̳Q̲ῐῲῐ΅̲̯Ῥ E. coli Ῑ῍ ῗ̰Ῡ̲ΌῺ̱̲΅Q̯ῬQ̮ῡ̮Ῥ1ῑ3)ῌ ESBL ῧ Q̮Q̰Q Q̮ῤΰῦQ̯Q̮΅̳ῚQ῞ῠ ῧQ̯ῦQ̲ῤ̱ ῐ´ ῒTEM-, SHV-, CTX-M Qΐ ῗΎ῍ ΊῚῤQQῺ῝ῢῤ̯ῠ̮Ῥ2)ῌ ῦQQῧ ῥ ῝῭ῠῗΎ “CTX-M pandemic” ῢQῨ῭ῠῗΎ῍ 2006 Ῡῦ In- 1ῑ3) ῍ 1990 ῩῚ̰ῡῧ῍ Qῲ΅̯ Q ῥ TEM- Q ῗ ̰ Ῡ SHV- Q ESBL ΅ Q ῎ ̯ Ῥ Es- fectious Diseases Society of America ῦ̱Qῡῧ῍ cherichia coli ̰ Klebsiella sp. Ῑ῾῝῭ῠ̮̯Ῑ῍ Q῞̮Q ̱ῦQ Ῑ̮QῥΎ̲ῢ῝῭Ῥ´QῙQQ 2000 Ῡ Q CTX-M QῙ̲ῐ ῶΏ΅̯Qῥ῾῝ ῦ̳̯ῥ῍ ῴQῙ̱ῷῶ´῝῭ῠ̮Ῥ4)ῌ ̯Q ̲ΐ῏ (̮461῍8673) ῷQQQῢQΊQῢ 1῍1῍20 ῷQQΊQ̳Q῭̱QQQ῎QQQQQQ Q ῖQ̰Q TEL: 052῍719῍3116 FAX: 052῍719῍1506 E-mail: [email protected] Q b-lactamase ῧ῍ CTX-M- 1 ̱ ῐ ´ ῥ ̮ ̯ Ῥ QQ῍ ̲ῐ ῶΏ΅̯QῥQQ῞ῠ̮Ῥ CTX-M-15 blaCTX-M-3 ῦ 240 ̰Όῦ̲ΌQῙ̱ (Aps̮Gly) ῞̯ῢῥ̰Ύ῎῟̯ b-lactamase ῡ῍ ̯ῦ ῡQῧ ´̱ Q IncFII ῦQΊ´῾̱̲Ὸῥ̰Ύῡ̰῞ ῠ̮Ῥ5, 6)ῌ ῦ CTX-M-15 Q b-lactamase ΅Q῎̯ Ῥ E. coli ῧ῍ ̯ῦ´῾̱̲ῸQῥ TEM-1, OXA-1 ῦ ῦῴ Q̰῎`QQQ Vol. 21 No. 1 2011. 25 ̲̲ῤ̳῎ῠ 26 ῬῖῦῠῨ b-lactamase Ύ̱Q ΌQΏ̲ ῌῖQ dards Institute (CLSI) QQ 9) ῧQῡ̮̯Q̰ῶ ̯ῲΏῑ ̯`Q̰Q῍ ῞῭ῧῩ qnr ῦῥῨ̳Q̲Q QQ ῧῢQ̮῍ Ῑῷ QQΰῙQ̰Qῲ Q̯`῝ῤῪ̮΅5, 6)῍ ̱ (CAZ), Q ΌQΏ̲ Q῍ QQ ̳QQ῍ ̳QῴQῷ Q SMX), gentamicin (GM), imipenem (IPM), levo- QQῦῥ̲ῘῨ̳Qῧῢ῟ῢΰῙῲQ΅῟ῢ̮`ῌ Q floxacin (LVFX), fosfomycin (FOM) ΐKB ̱̲Ὸ῍ ̮ΰῩ῍ ̱̲̱ῧῗ`̳QῨ῾QῩQῑQ ῧΌ QQQQ ῧῢ̯`QQῙῲ̰̰̯ῌ ̰̯῍ ESBL Q ̰ῡΊῩῦ̮Ῑ῍ ̱Q ῨQQQ̯Ῐ῭Ῡ̲ῷῧ῾Q ΊῙῨQῳῩ῍ CLSI QQ 9) ῧῒ̯̯ double-disk cefotaxime (CTX), ceftazidime trimethoprim/sulfamethoxazole (TMP/ 7) ῞ῢῗ΅ ῍ ̱̲̱ῧQ̰ῨQ´ῑ ῑῤῦ΅ῖ` synergy test ῗῬ̰ Etest ΐESBL CT/CTL, ESBL Q̱ῙῩῸ̰ΰῚῦ̮ῌ TZ/TZL, ̲̲QῸ̲῎ ῴQQQῑ ῧῢQ̯̯ῌ QQῨῤ῝῍ ̳ ΰῩ CTX-M-15 QΊ E. coli Ῠ 3. ESBL ῟̯̮ῡ̯῎Ὶ῞ῌῐ῏Ῑΰ ῾QῩ̰ῤῳῥ Q῞ῢ̮ῦ̮Ῑ῍ Q̱Ῠ̳QQQ ESBL QΊQ̰QῨQQῗῬ̰Q Ῡ῍ Shibata10) Ῑῼῑ̳̱QQ̲QQQῧῬ`̰Q Q ΐ2007ῒ ῗ Ῥ ̰ Yagi ῭11) Ῠ ῧ Ῥ ΅ PCR ῧ ῢ ῢ ̮ ̲ Q 2008 ̱ ΰῩ῍ ESBL QΊ E. coli ῨῡQΰῙQῲQ ΐCTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, SHV- ῗ Q̯` ῑ῾ῙQ῞ῢῗ΅῍ ̯Ῠ̯QῙQ̱῞ῢ Ῥ̰ TEM- Ῠ̯̲ῲQ̯̯Ῠ̯῍ ̯̲Q̲Ῠ̲ῑῸ ̮`ῌ QQ῍ QQQ̮ῧῗ`ῡQΰῙ ESBL QΊQ ̱Q̲Q῞ῲQ̯̯ῌ ESBL QΊQ̰QῨQῐῧQQ Ῠ̯QῤQQῲ Q̯`̯̰ῧῩ῍ ̰Q̰ῦ̳QQQ ῟̯̳Q̱̳ῑῲ Table 1 ῧQ̯ῌ 8) ῙῬQῨ̰QῤῤῪῧ῍ Qῐ῞`QQῨ`Ῑῧῠ̮ ῩQ DNA Ῡ Wizard Genomic DNA Purification ῢῪQ῞̯`῝ῤῙ῏Qΰ̮`ῌ ̯῝ΰ῍ ̳QQΰῩ Kit (Promega) ῲQ̮ῢῨῐ῟῍ ESBL QΊ E. coli Ῠ῾Q̲ΎῙQ̮ῲQQ̯̮QQ ̯ῌ PCR QῩῩQ DNA 1 ml (20 ng) ῤ PCR QQ ̱Ύ̰ῚQQQ῟ Qῲῢ̮ῧ῍ ESBL QΊQῨQQQῨQQῦ῭̰ῧ (1.0 U Taq polymerase, 0.2 mM dNTP mixture, 20 ESBL QΊQῤ̳QῴQῷ QQΰῙῤῨQQῲ̳῭ mM TrisῌHCl (pH 8.4), 50 mM KCl, 2.5 mM MgCl2 Ῐῧ̯`῝ῤῲ̳̰ῤ῟ῢ῍ ESBL QΊQ̰QῗῬ̰ (TaKaRa), 0.3 mM primer set) 24 ml ῲQQ῟῍ 95̮ 5 ̳Q̲Q Ῑῷ QQΰῙQ̰QῨ Q̮Qῲ̰Q̯ ῾QQ̱Q῍ ̱ Ῑ 95̮ 30 ´῍ ̱ ῑQQΌQῩ `ῤῤῪῧ῍ QΰῙQῧῠ̮ῢῩQ̰QQῨ῭̰ῗῬ Q̳Q̱̳ῑ̲ Ῠ̯̰QΎΰ 30 ´῍ ῗ̰Q ̰̯῭ΰῙQ̰Qῲ Q̯`̳Q̲Q Ῠ̰ῴῧῠ 72̮ 30 ´Ῠ 30 ῼ̱ῸQ῍ Q῍ῗ̰Q 72̮ 5 ῾Ῠ ̮QῧῢQ̯̯ῌ Q῍QQ῍ EtBr Q 2̮ ̱ῶQῑ ̮ῢῪQ῞ῲQ̯̯ῌ ̲ῺQῧῢῪQQῬῲQ̮῍ ̯̲Q̲ῨQ̳ῤῼ̱̲ ῲQῳ῟̯Q῍ QIAquick Gel Extraction Kit (QIA- ῝Ῠ῍ῤῥ 1. GEN) ῧῢ̯̲Q̲ῲ῝῟῍ ̲ῑῸ̱Q̲QῨῩ ̯ῧῘῖ QQῩ῍ 2007 ̱ 12 QῘ῭ 2008 ̱ 2 QῨ 3 QQ Qῤ῟̯ῌ ῧQῦQQ 7 Q̯ῧῗ̮ῢῲQQ̯̮QQῘ῭῾Q ̲ῑῸ̱Q̲QῩ BigDye Terminator v1.1 Cy- ῞̯ E. coli 312 QῲQQ῟̯ ΐQ῏ 151, QQ῏ cle Sequencing Kit (Applied Biosystems) ῲQ̮ῢ῍ 161, ῥῙ῏ 99, Ῑ῏ 213, Q̱Q 60.4 Qῌ QQῩ 95̮ 5 ῾QQ̱Q῍ 95̮ 30 ´῍ 50̮ 10 ´῍ 60̮ 4 QQ 104 CFU/ml Q̮ῲQ῟̯ῲQQ E. coli ΰ῍ ῭ ῾῍ 30 ῼ̱ῸQῨ̮QῧῢQ̮῍ ΅῭̯Q̲ῲ῝ QQQῧῠ̮ῢ῭̯Q̯̱̳̲ῑ ΰῨ῏̲ῩῺῢ Q ABI PRISMTM 310 Genetic Analyzer (Applied Q῎῟̯ῌ ̳QQQῙQQῨῼῒQQῤ῟ῢῩ῍ E. Biosystems) ῧῢQ῞῟̯ῌ ῦῗ῍ ̯῭ῙῨQ῞Ῡ῍ coli ATCC 29522 ΐ̱̱̳Q̱QQ ῲQQ῟῍ ESBL BLAST program (DDBJ, Shizuoka, Japan) ῲQ̮ῢ QΊQ̰Q QQῗῬ̰̳Q̲Q Ῑῷ QQΰῙQ Q̯̯ῌ ̰Q QQῨQQῧῗ`QῙΏQ QῑQQῩQQ 4. ̮ῒ̮ΐ̯ῑ̮̮ῠ̯̮ῡ̯῎Ὶ῞ Q̯̮QQΐῘ῭῾Q῞̯ῪῨῲQ̮̯ῌ ̰̯῍ ῟ ̳ Q ̲ Q Ῑ ῷ Q Q ΰ Ῑ Q ̰ Q ΰ ̮ ` qnrA, Q̰ῤQQῨQQQῤ῟ῢ῍ E. coli CSH-2 ῲ῍ Q̲ qnrB ῗῬ̰ qnrS ῧῠ̮ῢῩ῍ Cattoir ῭Ῠ 12) , ̳̱ῑQ ῺQῪQQῬῨΏQ QῑQQῤ῟ῢ E. aac(6̮)-Ib-cr ῧῠ̮ῢῩ Park ῭Ῠ 13) , qepA ῧῠ coli MC4100 ῦ῭̰ῧ Salmonella H9812 ῲQQ῟ ̮ῢῩ Yamane ῭Ῠ 14) ῧQῡῚ῍ QῐPCR ῧῬ ̯ῌ `QῐῲQ̯̯ῌ PCR QῩ῍ ̯ῑῨ ESBL QΊQ 2. ῦῗ̯̯̯Ίῌῐ῏ ESBL ῟̯̯῎̮ῢ ̳QQQῙQQῩ῍ Clinical and Laboratory Stan26 ̳QΏΊ̲QQQ Vol. 21 No. 1 2011. ̰QῨQῐ̮ ῤ῭QῧQ̮῍ ̱ ῑQQΌQῨQΎ Ῡ Table 1 ῧQ῟̯̯̰QΎΰQ̯̯ῌ QQ̱Q̳QQ ESBL QQQῶῚQ QQῗ CTX-M-27 ῚQ Table 1. Primers used in this study for PCR and nucleotide sequencing Annealing temperature (ῒ) PCR primers CTX-M-1-F CTX-M-1-R CTX-M-2-F CTX-M-2-R CTX-M-8-F CTX-M-8-R CTX-M-9-F CTX-M-9-R TEM-specific-F (T1)a TEM-specific-R (T2)a SHV-specific-F (S1)a SHV-specific-R (S2)a QnrA-F QnrA-R QnrB-F QnrB-R QnrS-F QnrS-R Aac(6῍)-Ib-cr-F Aac(6῍)-Ib-cr-R QeqA-F QepA-R 27 5῍-GCT GTT GTT AGG AAG TGT GC-3῍ 5῍-CCA TTG CCC GAG GTG AAG-3῍ 5῍-ACG CTA CCC CTG CTA TTT-3῍ 5῍-CCT TTC CGC CTT CTG CTC-3῍ 5῍-CGG ATG ATG CTA ATG ACA AC-3῍ 5῍-GTC AGA TTG CGA AGC GTC-3῍ 5῍-GCA GAT AAT ACG CAG GTG-3῍ 5῍-CGG CGT GGT GGT GTC TCT-3῍ 5῍-CCG TGT CGC CCT TAT TCC-3῍ 5῍-AGG CAC CTA TCT CAG CGA-3῍ 5῍-ATT TGT CGC TTC TTT ACT CGC-3῍ 5῍-TTT ATG GCG TTA CCT TTG ACC-3῍ 5῍-AGA GGA TTT CTC ACG CCA GG-3῍ 5῍-TGC CAG GCA CAG ATC TTG AC-3῍ 5῍-GGA ATA GAA ATT CGC CAC TG-3῍ 5῍-TTT GCT GTT CGC CAG TCG AA-3῍ 5῍-GCA AGT TCA TTG AAC AGG GT-3῍ 5῍-TCT AAA CCG TCG AGT TCG GCG-3῍ 5῍-TTG CGA TGC TCT ATG AGT GGC TA-3῍ 5῍-CTC GAA TGC CTG GCG TGT TT-3῍ 5῍-GCA GGT CCA GCA GCG GGT AG-3῍ 5῍-CTT CCT GCC CGA GTA TCG TG-3῍ Size of PCR product References (bp) 56.5 516 10) 53.5 779 10) 55.0 569 10) 53.5 393 10) 56.0 824 11) 55.0 1051 11) 54.0 580 12) 54.0 264 12) 54.0 428 12) 55.0 482 13) 60.0 199 14) Sequencing primer for TEM and SHVb T3 T4 T5 T6 S3 S4 S5 S6 a b 5῍-CAG TCA CAG AAA AGC AT-3῍ 5῍-CTG GCG AAC TAC TTA CTC-3῍ 5῍-TAG GCG GAG GTA GGT CAG-3῍ 5῍-TAC GAA AAG ACA CTG ACC-3῍ 5῍-GCG GGT GGA TGC CGG TG-3῍ 5῍-CGG CGG GCT GGT TTA TCG-3῍ 5῍-TAA ATC ACC ACA ATG CCC-3῍ 5῍-GTC GGC AAG GTG TTT TTC-3῍ 11) 11) To determine the genotype of ESBL, PCR was performed using the TEM- and SHV-specific primers (T1 and T2 for TEM. S1 and S2 for SHV). For nucleotide sequencing, the several sequencing primers (T3, T4, T5 and T6 for TEM; S3, S4, S5 and S6 for SHV) were used. 5. ῠῢῙ῏̯ῚῨ῍ῑῐ̮̮̮ΐῌ̮̮̮ ῥῘ̮ῦῩ῎ῑῒ̮ῤῗΰῧΊ῏ῖ̯ ̲̮̮ῌ 6. ̯῟ῤῡ̯῝῍ῑῐῤῡ῞̰῏̯̯ QQῶῚῺQῷQ ῐO ̲̲῍ H ̲̲ῑ Ί῍ QQῙ̳ QQQ ̲Ί῍ Qῶῗ̮̮ ESBL QQ̱̯῍ ̳ ΐQ̲QQῶQ΅ῺQ ῐῦῪ̯Q´ῑ ̯Qΐ̮ΌQ̮ Qῶῗ̮̮ E. coli CSH-2 ̱̯Qΐ̮ Broth Mating ̮ῌ Ῠ̰ῡῩ̯῎̰ῧ̯̰QῴῬQ (PFGE) Ί῍ Bar- Q16) Ῑ̮ῼQ̮̮ῌ LuriaῌBertani (LB) broth Ῑ̮Ύ rett ῝ῚQQ Ῑ̳ΐ Q̯QQ̮̮̲῍ XbaI Ῑ̮ QQQ̮̮ῶ῭̯QQῘ LB broth ῙQῸ̮῍ QQ Q̲῾Q̳Q̮῍ 6 V/cm, 2.2῏54.2 s Ὶ̳Ώῖ 19 QQ̱ (OD660: 0.5῏0.8) ῙῘ῞ῖ 37ῒῙ̮Qῗ ̱ ῐCHEF-DRIII ῠῡῥ̰῍ BIO-RADῑ QῴῬQ̯ QQ̮̮ῌ QQQQ̱ῙQ̮̮Qῶῗ̳Qῶ̯ 1.5 15) ̲̮̮ῌ QQ̲Q῝῟̮ PFGE QῙ̮ΐ̮Ί Finger- ml Q ῶ ̯ ῤ ̰ Ὺ ῧ ̰ Ῡ ΰ ̰ ῎ ̰ Q ῖ 1 : 4 Ὶ ̱ printing II (BIO RAD) ̯Qΐ̮̯̰ῡῢ῎ῲQ̯ ῐ100 ml ῗ 400 mlῑ Ῑ Q̮῍ 10,000 rpm 1 Q̱`Q QQQ̳QQQῳ Vol. 21 No. 1 2011. 27 ΰῢῐΎ῎Q 28 ῞̯Q῍ 37ῥ´῏ῷ̲ ῶῳῤῠ 20 QQ ῲ̲ ῗ 27 mm ̳̰῞Ῑῦ CAZ ῥ̯΅ῗ 22 mm ̳ ̲῏ῷ῞̯ῌ ῲ̲̲῏ῷQῥQ ῤQῺ Q῞̯Q ῑESBL Qῒ ῦ 40 Q̮Ῥ῍ ̯ῥ̮̯ dou- ῞῍ ̯ῥ̮̯ῥ 100 ml 20 mg/ml CTX ̮Ὺῧ 50 ble-disk synergy test ̮Ὺῧ Etest ῡ ESBL QQQ mg/ml rifampicin ̳̮̯ LB Q̮̯ῒῤQQ῞῍ ῢ̯̮῝΅̯Qῦ 15/312 Q (4.8̮) ῡ̮̯̯ῌ ΅ ̱῏ ῾῏̱̰ῡ Ὺῤ̮ῬQΊ̯Q῍ 37ῡ̯ῳ῞ Ύ 15 QῥQQ῟Q Table 3 ῤQ̯ῌ ῍Qῢ῞̯ ̯ῌ 48 QQQ῍ Q̮ῦ̯QῤῗΎ΅̯̱ Ό῏QQ 312 ῶῥῦQ῝ῷῗ 60.4 Qῡ̮῭ῥῤ῍῞῍ 15 ῶῥ ̮ῐῌῢ῞῍ ̳ῥ̳QQ̯̯ῌ ῦQ῝ῷῦ 72.8 Qῡ̮Ῥ῍ ESBL QQQῦQῷQῖ ῦ῟̰ῤ PFGE ῤῪῬ῍ QQ̮ῐῌ῍ QQῢQ ΎQῙῥ̱῝΅῭QQῗΊ̰Ύ΅̯ῌ Qῧῡῦ῍ ῑQ QῥΏ Ὸῼ῏ ̯Q῞῍ ῗΎ΅̯QQ̮ῐῌῗ 4/99 (4.04̮), QQ 11/213 (5.16̮) ῢ΅ ΰQῦΊ QQ`̱ E. coli ῡ̮῭ῢQΊ῞̯ῌ Qῤ῍ QQ ̰Ύ΅ΰῖ̯̯ῗ῍ Ῑ QQ`̱ῗ 8 Q῍ Q̱QQ` ̮ῐῌῥ̲῾̱̲Ὸ DNA QIAprep spin plasmid ̱ῗ 7 QῢῩ̰ῖQῥ̱῝΅ῠ̮Ῥ῍ QΐQQQῤ̮ kit (QIAGEN) ῤῠQ῞῍ Q῏̲῾̱̲Ὸῥ̮ῐQ Ὶ῭ ESBL QQ E. coli ῥQ̳QQῗ̰Ύῖῢΰ̯̯ῌ Q Ί ῞ ̯ῌ ESBL ῠ Q Q ̮ ῐ ῥ ΅ ̰ ῤ ̯ ̮ ῠ ῦ LVFX ῤ῍῞῎QQ῞̯ 49 Qῥ̮̯῍ PCR ῤῪ double-disk synergy test ̮Ὺῧ Etest ῤῠ῍ TMP/ ῭QQῡ̲῾̱̲ῸQῲῺ ῎Q ̮QῥῩ΅ῗQ SMX ῢ CPFX ῥQQQῥῨ̳ῤ̯̮ῠῦ Disk ̰ῤ Ί ῝ ΅ ̯ Q ῦ῍ qnrS ῲ Q ῥ 1 Q ῥ ̰ ῡ῍ ῥ Q ῦ ῠ̮Ῠ̯ῌ ESBL ̯QQQῡ̮̯̯ῌ ̰῍ ESBL QQQῡῦ῍ ΰ̮῍ QQ̮ῐ῞̯̲῾̱̲Ὸῥ̱ῴῦ῍ Q῞ 15 Qΐ 13 Qῗ LVFX ῤ῍῞῎Qῡ̮Ῥ῍ ῝Ύῤ῍ ̯̲῾̱̲Ὸ DNA QQQQ Pst I ῡQ̱῞̯Q῍ ῗ ΅Ύ LVFX ῎Q 13 Qῥ̮̯῍ 7 Qῦ TMP/SMX Ύ΅̯Ώ Ὸῥ̱ῴῖΎQQ῞̯ῌ ̰̯῍ ̮ῐQ̱ ῤ῍῞ῠ̰῎QQῗ῞ῠ̮Ῥ῍ ESBL QQQῤ̮Ὶ ῥQQῦ῍ QQQQῤ̮Ὶ῭QQῥ̱ Ό῏Qῖ ῭QQ῎Q̳QQῗΊ̰Ύ΅̯ (Table 3)ῌ QῤQ̮῞῍ QQ̮ῐ̱ Ό῏Q / QQῥ̱ Ό῏ Qῢ῞ῠQQ῞̯ῌ ESBL Q Q 15 Q ῥ ̮ Q Q ῧ Q ̯ ̯ Q ̳῍ blaCTX-M-1 ̮Ὺῧ blaCTX-M-2 Ῡ΅QῗQ 1 Q῍ CTX-M9 ̱ ῏ ̲ ῥ blaCTX-M Ῡ ΅ Q ῗ 12 Q ῑblaCTX-M-14; ῍ ῌ 4 Q῍ blaCTX-M-27; 8 Qῒ῍ blaTEM-1 Ῡ ΅ Q ῗ 3 Q῍ ῚΌQQQ`̱ E. coli 312 Qῤ῍̯῭῭QQQQ blaSHV-12 Ῡ΅Qῗ 1 Qῡ̮Ῥ῍ blaCTX-M-14 Ῡ΅Qῥ̮ Q Q Q Q ῞ ̯ Q ̳῍ 7 ῭ Q ῤ ῍ ̯ ῭ ῎ Q ̱ ῦ῍ ̯ 3 Qῗ blaTEM-1 ̰Ῡ΅῞ῠ̮̯ῌ ̮QQῧῢ῭Q LVFX (15.7̮), TMP/SMX (9.6̮), GM (9.3̮), CTX QQQQQQ῭ῢ῍ CTX-M-9 ̱῏̲ῥ 12 Qῦ (4.8̮) ῥQῤQῙ῍ CAZ ̰ FOM ῤ῎QQ̯Qῦ ̯Ῠῠ LVFX ῎Qῡ̮Ῥ῍ b-῾ῳ̲QQQ῭ῤ̯ 1̮ ̳῍ ̰̯῍ IPM ῤ῍῞ῠ῎QQ̯Qῦ 1 Q̰ ̮ῠῦ῍ blaCTX-M-14 Ῡ΅QῪῬ̰ blaCTX-M-27 Ῡ΅Qῥ Ί̰Ύ΅ΰῖ̯̯ῌ ΅Ύ῎Q̱Ῑ ̮ῪῧQ̱Q Ῡ ̮ ῗ CAZ ῤ ῍ ῞῍ Ὺ Ῥ ̮ Q Q Q Q ῞ ῠ ̮ ̯ Q`̱ῧῡ̯Q̯῭ῢ῍ LVFX ῤ̮̮ῠ῍ Q̱QQ` (Table 3)ῌ ̱QῥῩ̮ῗ̰̰Q̮QQῗΊ̰Ύ΅̯̰ῥῥ῍ ̯ῥ QQQῧῡῦQQQQQ῞̯̮ῡ῞ῥ̰ῥῗ 6 Qῥ῭Qῡῦ̱QῥQῤ῏ῘΰQῦΊ̰Ύ΅ΰῖ̯̯ Q̮̯̯ῗ῍ CTX-M-9 ̱῏̲ῡῦQQQ O25 : H4 (Table 2)ῌ ΅Ύ 312 Qῥ̮̯῍ CTX disk ῥQQ̳ ῗQῙΊ̰Ύ΅̯ (Table 3)ῌ PFGE ῤῪ῭ ̮QQ ῧῡῦ῍ QQ A ῡ 3 Q (No. 41, 45, 79), QQ C ῡῦ 4 Table 2. Antimicrobial susceptibility of 312 Escherichia coli stains Antimicrobial agents CTX CAZ IPM LVFX GM TMP/SMX FOM Number of resistant isolates Total 15 2 0 49 29 30 1 Inpatient Outpatient ( 4.8̮) 8 (2.6̮) 7 (2.2̮) ( 0.6̮) 0 (0.0̮) 2 (0.6̮) ( 0.0̮) 0 (0.0̮) 0 (0.0̮) (15.7̮) 19 (6.1̮) 30 (9.6̮) ( 9.3̮) 13 (4.2̮) 16 (5.1̮) ( 9.6̮) 12 (3.8̮) 18 (5.8̮) ( 0.3̮) 1 (0.3̮) 0 (0.0̮) 28 Ῐ̰ῴQ̯QῤQQQ Vol. 21 No. 1 2011. Q (No. 22, 70, 179, 186) ῡQQῗΊ̰Ύ΅̯̰ῥ ῥ῍ QQQῤQ̮QῸQῦΊ̰Ύ΅ΰῖ̯̯ῌ ΅Ύ 7 QῥQQQῢ`̱Q῭ῢ῍ QQ No. 22 ῢ 41 Q Ῑ 5 QῗQQQ O25 : H4 ῡ̮Ῥ῍ QQ A ῥ 3 Qΐ 2 Q (No. 45, 79) ῢ῍ QQ C ῥ 4 Qΐ 3 Q (No. 22, 179, 186) ῗQ̱QQ`̱Qῡ̮̯̯ (Table 3, Fig. 1)ῌ ESBL QQ 15 Qῤ̯̮ῠ῍ Broth Mating ̰ῤῠQ Q̮ῐQQQQ῞̯ῢ`῍ CTX-M-9 ̱῏̲ῥ ̮̯῍ 4 Q (No. 70, 79, 179, 239) ῤ̮̮ῠQQ̮ῐQ ῗQΊ῝΅῍ ̯΅Ύ̲῾̱̲Ὸ̱ῴῦῬ 40ῐ50 kbp, QQ̮ῐQ̱ῦῬ 3.410ΐ5 cells/recipient ῡ ῪQQQῠῧ ESBL QQQQQΰQQQQ ̯ CTX-M-27 ΰQ Table 3. Genetic and phenotypic traits of ESBL-producing Escherichia coli strains Inpatient/ Strain Sex Age number (M/F)a Outpatient 10 22 23 25 41 45 70 79 81 147 179 186 211 239 242 a b c d e 86 90 64 70 85 79 79 94 23 43 70 61 70 84 94 F M F F F M M M F F F F F F F 29 Inpatient Outpatient Outpatient Inpatient Inpatient Outpatient Inpatient Outpatient Inpatient Outpatient Outpatient Outpatient Inpatient Inpatient Inpatient ESBL genotypeb CTX-M-2 CTX-M-27 CTX-M-27 CTX-M-27 CTX-M14, TEM-1 CTX-M-1 CTX-M-27 CTX-M-14, TEM-1 CTX-M-14 SHV-12 CTX-M-27 CTX-M-27 CTX-M14, TEM-1 CTX-M-27 CTX-M-27 MIC (mg/ml)c Resistance phenotype CTX CAZ to non-b-lactamd 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 16ῒ 0.75 4 8 4 1 32ῒ 2 0.5ΐ 0.5ΐ 0.5 1 2 0.5 2 2 LVFX, LVFX, LVFX LVFX LVFX LVFX, LVFX LVFX, TMP/SMX TMP/SMX TMP/SMX TMP/SMX, GM LVFX, TMP/SMX LVFX, TMP/SMX LVFX LVFX, TMP/SMX LVFX Serotypee Hospital O125 : H4 OUT : H4 OUT : H4 O25 : H4 OUT : H4 O25 : H4 O25 : H4 O25 : H4 O169 : HUT OUT : HUT O25 : H4 O25 : H4 OUT : H4 OUT : H4 O25 : H4 B C C C A A C A E B C C D B B M, male; F, female b-lactamase genes were determined by PCR and by sequencing. MICs were deterimined using E test “ESBL CT/CTL” and “ESBL TZ/TZL.” LVFX, levofloxacin; TMP/SMX, trimethoprim/sulfamethoxazole; GM, gentamicin OUT, O-antigen untypeable; HUT, H-antigen untypeable ̯῞ (Fig. 2)ῌ ῎̰Ὺ῞QQQQQ 4 Qΰ̯QQQ QQ ῐ4.8, 15/312 Qῑ ῤ῍ 2006 ̮ΰ Yamaguchi QΎQ̰̯῍ QῢQ E. coli CSH-2 ΰ LVFX ̮̰ῥ ̰ΰQ ῐ4.3, 6/140 Qῑ21) ̰ 2009 ̮ΰQQ̰ΰ GM QQQῤ̲QῚῪ̯̮῞̮῍ CTX ῢQΊ̯ῤQ Q ῐ6.3, 13/205 Qῑ8) ̯ῦῧ῍QQ̯̰῍ Ῑ QΎQ῎Ί̯̮̰῍ double-disk synergy test ΰQ̳ ̯QῚῪ̯̮̰ῦῠΰQQῡQ ῤῩ̰Ὺῡ̮̯ ESBL QQQ̯̮QῚῪ῞ῌ Ὶ̰ῢ῍ QQQQQ 3 Q ῞ῌ Ί̮Ίῡ̮̰῍ ESBL QQ 15 Qΰ̮῟῍ 86.7 ῤ TMP/SMX ῢQΊ̯̰QῡQ̯῍QQΰQQΎQ ῐ13/15 Qῑ ̮ LVFX ῢQΊ῍ 40.0 ῐ6/15 Qῑ ̮ Ί̯̮῞ῌ LVFX ̮̰ῥ TMP/SMX ῢQΊ̯QQΎQ῎Ί̯̮ ̰῍ Ῑ̯῍ΰ῍ ESBL QQQ̮QQQQ Ί̯̮̰ ῌ ῍ Ῑ̯̮῟̰̮̯ῡ̯῞ῌ Ῠ῞῍ Q̳ῗῩῚῪ῞ ESBL QQ῍ E. coli ΎQ Q̯̰QQQQQῪQQQ QQQQQῢ̯̮̯QQ̮QΎQ̰̯῍ Q̰Q̮̰Q ΰQQQ̯̯Ί̯῍ QQ̱ῶῬ̲̱Όῼ̲QQQ ῖῗῩῚῪ̰QQῤῙῪῨ̯ΰQ2) ̯῍ΰ̯̯῞ ̯῍ ῶ´̲̲̲QQQ̯῍ TMP/SMX ΰQῥ̮ ̮῍ ̳ῧQQῠῧQ̯ΐ QQῠῧQ̮ῦῧ῍QῗῩ QQῚῪ̯̮̰ ῌ Ί̮Ίῡ̮̰῍ E. coli ΰ̯ 15῏ ῚῪ̯̮̰῍ ESBL QQ E. coli ̮QQQQQΰQ Q 20 ̮ TMP/SMX ῢQΊ῍ ̯ 10῏25 ̮ῶ´̲ ̯Ί̯̯Q̯ῡ̰̯̯̰QQΎῨ̯ῗ̰Q̳̯ῡ̯ ̲̲QQQ̯ῢQΊQQΎQ῎Ί̯̮̰̯ΰQ ῞ῌ Ῑΰ ESBL QQQΰQQQQ QQ̯̳ῧQQῢ ̰̰18, 19)῍ ῪQQQQ̰ῢ̮ῗ̰QQ̱ῶῬ̲̱ ̮ῗ̰Q QQῤ῍ ῪQQQῢ̮ῗ̰QQ̯Q̰Ύ Όῼ̲QQQ̯ΰQῦ̮QῨ̰̯̯̰ῌ ῝ΰ῝ Qῑῢ̰ ̮QΎQQΊ̯̮̰̯̰ῚῪ̰ῌ 17) ̯῍ Ῑ̰῭̱῭QῘῤQ῾ῡῠQ̳̳Qῢ̮ῗ̰Q CTX-M Q ESBL QQQῢ̮ῗ̰ῶ´̲̲̲ Q̯ῳ῎Ὸ΅Ώ̲̱ῢ̰̰῍ E. coli ̰ K. pneumoniae QQ῍ QQQῡ̰ῥῢ̱῎ῲ`̲̱̱΅ῷ ῐ῏ῢ ST ῡῠΰQῐQQ QQῢ̮ῗ̰ ESBL QQQΰQQ 131ῑ ΰQῢῤ῍ QΰQ̰Q̮Ῡ̰Ὺ̰̯ΰQ ῡQ QQ̮῟̰̮̯ῡ̰6, 20)῍ ̰QQQῢ̮ῗ̰Q ̮̰5, 22)ῌ ῙῪ̰QQῢ̮ῗ̰ῶ´̲̲̲Q Q̯QῥῢQ̮ῡ̯QΎῌῘ̮ῗ̯̮̰ῌ Q̳ΰQQ QΰQῖῤ῍ ̲̲QQQQ̰῾ (QRDR) ΰQῚ̲ ̯QῥΊ῞ 2007῏2008 ̮῍ ̲QQ 7 QQ̯ῗῩῚ ῢ̰̰̮ῡῚῪ̯̮̰̮῍ ῷΏ̱ῺῴQ̲̲QQ Ὺ῞ῪQQQῠῧ E. coli ῢ̮ῗ̰ ESBL QQQΰ ̲QQΎΊ̯̰Q̰Q̯̮̯̯̮̰5, 22)ῌ Q̳̳Q ῒ῞̰Q̮QῖQQQ Vol. 21 No. 1 2011. 29 Fig. 1. Conjugation of plasmid and susceptibility profile of resulted transconjugant Transconjugation analysis was performed with E. coli CSH-2 as the recipient by the broth mating method. Transconjugants were selected on LB agar plates supplemented with rifampicin (50 mg/ml) and CTX (20 mg/ml). Plasmid DNA from recipient, donor, and the resulted transconjugant were purified using QIAprep spin plasmid kit (QIAGEN). Figure shows those of No. 179 among four isolates subjected to the transconjugation analysis. The susceptibility profiles of recipient, donor, and the resulted transconjugant were also shown. ESBL phenotypic tests were performed using E test (SYSMEX bioMe ´rieux). The susceptibilities of LVFX and TMP/SMX were determined by agar di#usion method according to CLSI recommendation, and categorized into susceptible (S), intermediate (I) and resistant (R) in accordance with CLSI criteria. 30 30 ̮ῖῑ̮ῒ̮῎῏ῐ ̮̮̮ῌΐ Vol. 21 No. 1 2011. Q̮̳QQῑ ESBL QQQQQ̯QQQQ ̯ CTX-M-27 ̯Q 31 Ῑ CTX-M Q ESBL Q Q Q ῠ Q ̳ LVFX Q Q ̯ ῞̮῍ Ί̰̰̯Q̯ῶΌῳῷ̱Q῭̱ῼ´QQ̲ QQQῐ̳ῠQῦ̰̰̯̮῞ῌ ῾Q῍ ESBL QQQ ̯QQQῠ O25 : H4 ̮̮῝QQQ̮̰Qῦ̯̮̰῍ ῗ̰̰ῠῚῢ̯ LVFX QQ̯῞ῌ Q ῠ ST Q Q̰Q῞̯̯̯̯῍ ῗ̰̰̮QQῸ῎ῼ̱̱̯Q Ῑ̯̰ ST131 ̯̰̮ῠQῌ̯̰̮῍ Q ̯ Q ῠQQQ O25 : H4 ̮ῴΏῬῼ῭̱ῼ´QQ̰ QῙῨῚ̯̮ Q̯̯QQ5, 22) ̯῾QῙ̯̮̰῍ QQ̯̮̯ῧ O25 : H4 Q̯ῴΏῬῼ῭̱ῼ´QQ ̰ QῙ ESBL QQ E. coli ̮Q Ῑ῟῟̰ῗ̯ ̮QQῘ̰ῌ QQ῍ ESBL QQQ̮ῶΌῳῷ̱Q῭ ̱ῼ´QQ̲QQ̰QῐῙ̯̰̯̯QQῧ̰22)῍ QQῠQῐQQ̯̰ῡ̯QQQQ̯῟̯ῧQ̲̰Q ̯̮̰῍ QQ̯QQ̰̳QῚ̰Q̮̰̯Q̰̰ ̰ῌ ESBL ̯Q̯`Ώ῎ῶ̯̮῝῍ Ὸ῎ῼ̱̱̯ῠ CTX-M-15, ST131 (O25 : H4) Q̮̯ E. coli ̮῍ Ὺῲ Fig. 2. Dendrogram generated by Pulse-field gel electrophoresis For molecular typing, the chromosomal DNA of fifteen ESBL-producing isolates was obtained for comparison by Pulse-field gel electrophoresis (PFGE) using a CHEF-DRIII (BIO-RAD, Tokyo, Japan). The running parameters were as follows: initial pulse 2.2 s, final pulse 54.2 s, at 6 V/cm for 19 h at 14ῒ. The gel was stained with ethidium bromide and scanned. Both E. coli MC4100 and Salmonella H9812 were used as a DNA reference standard. The DNA pattern obtained by PFGE was analyzed with Fingerprinting II (BIO RAD). The unweighted pair group method (UPGMA) was used to generate a dendrogram with the position tolerances set at 1.0ΐ, and the Dice coe$cients were used for band similarity measurements to determine DNA relatedness. Isolates were considered to be clonally related if the Dice coe$cient correlation was over 80ΐ, which corresponds to the “possibly related” category according to the criteria suggested by Tenover et al.26) OUT, outpatient; IN, inpatient. ῪQQΐQ̲̯ῠ CTX-M-9 `Ώ῎ῶ̯ E. coli ̮῏ ̲ ̯ ̰ ̯ Q Q Ῐ ̰ ̯ ̰1, 2, 16)ῌ Q Q ̯ ̮ ῖ ̰ ESBL QQ E. coli ̯῟̯ῠ῍ 1997῏1998 Qῥ̯ῠ CTX-M-2 `Ώ῎ῶ̮Q̮̯῞̮῍ 2000 QQQ̮ ̰ CTX-M-9 `Ώ῎ῶ̯Q̰Q̰῞ῗ̯̮ Suzuki ̰̯QQ̯Ῡ̰ῌ̰̮̯Ῐ̰̯̰23)ῌ Q ̯QQ̰ ̯QQ̯ῠ῍ CTX-M-9 `Ώ῎ῶ̯ ESBL QQ̳̯̳ Qῠ 73ΐ ῐ11/15 ̳ῑ ̯῍ Suzuki ̰̯QQ ῐ65ΐ ῐ84/142 ̳ῑῑ23) ̯ΰῤ῾QῙ̯̮῍ Ί̯̱ Ύῶ̰Q Ὶ̰̯ Suzuki ̰̮QQQQ̯Ῑ 2002 ῏2003 Q̯ῠ῍ blaCTX-M-9 ̯ blaCTX-M-14 ̮Q̮̯῞ ̯̯QῙ῍ Q ῠ blaCTX-M-27 ̯ blaCTX-M-14 ̮Q̮Q ῦ̰̰῍ ̲QQQ̮Q Ῑ῟῟̰ῗ̯̮ῌ̰̮̯ ̯῞ῌ blaCTX-M-27 ῠ blaCTX-M-14 ̯ 240 Q῍̯Ὺῷ̱ Q̮Ὺῳ̱Ό΅´Q̮̰`Ὼ´̯Q̲Ῑῧ̯̯ ̰῍ ῗ̯Ὺῷ̱QQ̲̮ CAZ QQ̯QῒῚ̰ῗ̯̮ QῘ̰̯̰24)ῌ ῗ̯ῗ̯ῠ blaCTX-M-27 Qῐ̳̯ΰ̮ ̮ CAZ ̯Q̳QQ̰QῙQQ ̯῾QῚ̰ῌ QQ ̯̮̯῍ CAZ ̯Ῡ̰QQ̰QῚQQQ O25 : H4 ̯ blaCTX-M-27 Qῐ̳̮ CTX-M-9 `Ώ῎ῶ̯QQ̲Q̰ Qῦ̰Ῡ̮̯̯῞̯̮̯̮̰῍ Ῡ̰῞Ί̮̯Q̲̮ Q̯̰ῌ ESBL QQ E. coli 15 ̳̯̮῝῍ ῎QQ̮QQQ O25 : H4 ̯̰῍ Ί̰̰ῠQQ A (No. 45, 79), QQ C (No. 70, 179, 186) ̯QQῙ̯ῌ PFGE ̯Ῡ̰̲Q QQQ̯ῠ῍ Q῾QQQ ῐA ̮Ῡῡ Cῑ ̯̮QQ̮Q ῦ̰̰ῧ̯̯῍ QQ A ̯ 3 ̳Q 2 ̳̯QQ C ̯ 4 ̳Q 3 ̳ῠ ̳Qῑ̳̯῞ῗ̯̮̰῍ ῗ̰̰ QQ̮QQQQ̳QQ Vol. 21 No. 1 2011. 31 Ὸῷ̯ ῏῝ 32 ῤῴ Qῗῢ̰̰Q̮̯̯Ῡΰ̯ῡ̮ῠQῪ̰̰ῌ ῍̮῍ ̲QQῴ ῍ QQ̳Q ῴ ῍ QQ Suzuki ̰ῤ῍ 2000 ῲ̲Q῍ CTX-M-9 ῼῐῷῢῚ ῴ ῍ Q̳̰ῴ ῍ Q̳ῦQ῾Q̱̲̱ῐ῍ ̲QQ ῞ ̰ Q ΐ Q O 25-ST 131 ῠ O 86-ST 38 ̮ ̲ ̲ ΰ ῴ ῍ Q QQ̳ῴ ΰ̱ΌQQQΰ̳̳ῢQ ESBL Q E. coli ΰQQ̯̰ῙῠΎ̲Q῍ ̯̰̰ ̰̰QQῌQῘῧ῞ῌ ΰ̲̰QQΏῢ̯̮̯Ῡ̳̮̯̮̰ ῌ ̯̰ῢ̰̰ 23) ῏ ῠ῍ ̰῝ O25 : H4-ST131 ῢῚ̯̮̯Ῡ῍ PFGE ̱ῐ̲̮̱῟Ῡ̰̯ῡ̮Ῑῠ̮QῚ̰̯̮̰ῌ Q 1) ̳῍ 2 Q̮ΰ` ̮̰ῥ̳̳QQ ̳Q̮̰῍ ̲̰Q QΏ̮QQ̯QΐQ O25 : H4 ̮QQῚ̰̯Ῑῠῤ῍ Ὼῐ´̱ῠ̰̳ῢ῍ ΎῨΰ´ῐ̲ΰQ̮̰ΎQQ 2) ῞̰Ῡΰῠ̯QQ̮ῌ QQῤ῍ ῝ΰQQQQ̮̰ Ὼ̳Ὶ̰̰ ESBL QQῩ῟Qῠ῍ ST QΏΎQῨ ̯QQῡῺQ QῩῡ̳̮ΎQ̮῍ ̯ΰ̰QΎQQῢ 3) QQ῞̰̱̳̮̰ῠQῪ̰̰ῌ ESBL Q̲̰Qῤ̮Q̰̯ῑΎῩ̯Q̯ῷΏῳΌ ̱̯̰̱῞̰Ῑῠ̮῍ QQΰ῾ ΰQ ΰ̯ῠῡ̯ 4) ̯̮̰ῌ Q̳ῥῦ̯ ESBL QQῢ̯̮̯Ῡ῍ 15 Q ΰ 4 Q̯̮Q̰̯ῑ̮QῚ̰῍ Ὺ̰̰̯̮Q̰̯Q 3 Qῢῤ῍ TMP/SMX ̯ῑῩ̰̯Ὶ̰̯̮̯ῌ TMP/ SMX ̯ῑῢ̯̮̯ῤ῍ blaCTX-M ´ ῷΏῳΌ̱Qΰ Ώῳ 1 ῭̲̱´̲Ῥῢ dfrA ̮Q῞̰Ῑῠ̮̲ 5) QῚ̰̯̮̰25)῍ TMP/SMX ̯ῑ̮̰̯Ὶ̰̯ 3 Q ΰῷΏῳΌ̱QῢῩῙΰ῭̲̱´̲̮Q῞̰̳ῳ ῑ̮QQῚ̰̯ῌ QQῗQῤῴ Qῗΰ ̮Qῢῡ ̰ῙῠῩ῞ῖ῍ ̯ΰQ Qῢ̮Q̰̯ῑΎ ῞̰ῷΏ 6) ῳΌ̱̮Qῶ̰ῢQῚ̰̯Ῑῠ̮̰῍ ῾QQQῢ̮ ῗ̰ ESBL QQΰῚ̰ῡ̰QQ̮QQῚ̰̰ῌ QῴQῑ̯ῤQΰ 5̮ ESBL QQ ῒQῢ CTX-M Qΐ ̯̯̯ῌ ̯̰̰ῤῠQῑQQῗQΰ 7) ̯Ῐ̯ ̯̰ῶῼ΅´`ῴ´̲QQQ ̰ TMP/SMX ῢ῟̯Ῡ̯ῑΎQῪ̯̮̰῍ ῞Q̯ ῑ QQῢ̰Ῑῠ̮ ̰̮ῠῡ̯̯ῌ Q̳῍ ST Q ΏΎQQ̯̮̰῟QQQΰQΎΎῨ̯̮ῡ̮̯̯ 8) Ῡΰΰ῍ O25 : H4 Qΰῶῼ΅´`ῴ´̲̯ῑ ῍ blaCTX-M-27 ´ Qῢ̮ῗ̰ CAZ ῧQQῑ ̮QῚ ̰῍ ῧ̯ CTX-M Q̲̰QΎ´ ̯ῷΏῳΌ̱Ῡ 9) Qῶ̰̯̰̱῞̰Ῑῠ̮ ̰̮ῠῡ̯̯ῌ ESBL Q Qῤ῍ ῷΏῳΌ̱Qΰ῭̲̱´̲ῢῚῧΊῧῡ̯ῑ ̲̰QΎQ̮̯̯῍ Ὶ̰ῢ̯ῑΰ̱̲ΎQῘ̰̲Q 10) ῢῬΌῴQῼ̲Ύῢ̮̯῎ Ίῗ̯̮̰ῌ Ῑ̰̰ ῞Q̯ῑ ESBL QQΰ῾ ΎQ̮QῨ̰̯Ῠῢῤ῍ ̯ῑQΰῺQ QῩῡ̳̮̮̰ῥ̮QῩῡ QῥQ̮ ̰῍ ̯ΰ̱QΎῡ̰Ῑῠ̮Q̳̯̰ῠQ̮̰ῌ ῎ ῍ ̲QQΎῐQ῞̰ῢ̯̰῍ QQΰῺ̳Ύ Q̰ῧ̯Q̳QῗQQQQQQ̯῭ῷῷῦ QῖQ 32 ΅̲QQ̱ΌQQQ Vol. 21 No. 1 2011. 11) ῌ Bonnet, R. 2004. 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Microbiol. 33: 2233ῌ2239. ̰῭ῠΎῤῬῚ̯̯ Vol. 21 No. 1 2011. 33 ̮̮ῖῌ 34 Multidrug-Resistant Phenotype and Prevalence of CTX-M-27 in ExtendedSpectrum b-Lactamase-Producing Uropathogenic Escherichia coli Tatsuya Hattori,1) Kunihiko Nakane,1, 2) Kumiko Kawamura1, 3) 1) 2) 3) Department of Pathophysiological Laboratory Science, Nagoya University Graduate School of Medicine City of Okazaki Reseach Center, Hygiene Testing Group Department of Medical Technology, Nagoya University School of Health Science The susceptibilities against 312 uropathogenic Escherichia coli strains isolated from clinical specimens were investigated, and 15 were extended-spectrum b-lactamase (ESBL)-producing isolates. Among the 15 ESBL-producing isolates, 13 were resistant to levofloxacin (LVFX) and 6 were resistant to both LVFX and trimethoprim/sulfamethoxazole (TMP/SMX), suggesting that ESBL-producing isolates tend to become multidrug-resistant (MDR) phenotypes. The genotypes of ESBL were determined by PCR and sequencing. Four isolates were CTX-M-14 producers and 8 isolates harbored blaCTX-M-27, which harbor the Asp240Gly substitution, have greater catalytic e$ciencies against ceftazidime than parental enzymes. None of these isolates harbored plasmid-mediated quinolone resistance genes, but 6 were serotype O25 : H4 among 13 ESBL-producing and LVFX-resistant isolates, suggesting fluoroquinolone-resistant phenotypes are associated with serotype O25. The CTX-M encoding plasmid was successfully transferred to recipient E. coli CSH-2 at a frequency of 3.4῎10῍5 cells per recipient by conjugation in vitro, suggesting the easy dissemination of blaCTX-M-harboring plasmids. Moreover, the transconjugants obtained showed both ESBL-producing and TMP/SMX-resistant phenotypes. Among uropathogenic E. coli, the serotype O25 : H4 isolates harboring CTX-M-27 blaCTX-M have become prevalent strains, so due care should be taken in monitoring the epidemiology of ESBL-producing isolates. 34 ̮̮ῗῒ̮ΐ̮῏ῐῑ Vol. 21 No. 1 2011.
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