Klebsiella pneumoniae 21 PCR

Transcription

1 TEM-132 ESBL Klebsiella pneumoniae 1) 2) 1) 1) 3) 2) 1) 2) 3) ceftazidime (CAZ) Klebsiella pneumoniae 21 PCR b- (ESBL) PCR (PFGE) PCR bla TEM-132 PFGE 19 TEM-132 b- K. pneumoniae CAZ- K. pneumoniae ESBL SHV b- TEM-132 b- Key words: TEM-132 ESBL Klebsiella pneumoiae I. Klebsiella pneumoniae Escherichia coli b- (extended-spectrum b- lactamase: ESBL) K. pneumonia E. coli 1), 2) ESBL 3) ( ) 1 23 TEL: FAX: kaetsu saikin@niigata-min.or.jp K. pneumoniae ) ESBL 5), 6) ceftazidime (CAZ) K. pneumoniae K. pneumoniae CAZ- K. pneumoniae 25 CAZ- K. pneumonae (PFGE) II Vol. 21 No

2 12 1. ESBL MIC (mg/ml) MIC 90 Ampicillin Piperacillin Cefazolin 4 8 Cefotiam 1 1 Ceftazidime Aztreonam Latamoxef 8 8 Imipenem Gentamicin 2 2 Minocycline 4 4 Fosfomycin Tosufloxacin K. pneumoniae, 378 BBL CRYSTAL E/NF 5 Sheep Blood Agar 1ml (HI) (McFarland) No. 0.5 Gram-negative 10 ml MICS ml ampicillin (ABPC), piperacillin (PIPC), cefazolin (CEZ), cefotiam (CTM), latamoxef (LMOX), CAZ, aztreonam (AZT), imipenem (IPM), gentamicin (GM), minocycline (MINO), fosfomycin (FOM), tosufloxacin (TFLX) 12 (CLSI) 1 2. ESBL ESBL ESBL Confirmation Panel CLSI CAZ CTX (CVA) MIC 3 ESBL Double disk di#usion test 3. ESBL PCR PCR ESBL PCR Shibata 7) TEM- b- TEM- b- TEM- b- (TEM-SQ-F: 5 - ATAAAATTCTTGAAGACGAAA-3, TEM-SQ-R: 5 -GACAGTTACCAATGCTTAATCA-3 ) PCR PCR 4. (PFGE) DNA XbaI PFGE Gene Pulser: V/cm, ) PFGE Tenover 8) 5. ESBL PCR, PFGE III. 1. BBL CRYSTAL E/NF K. pneumoniae 2. ESBL ESBL K. pneumoniae MIC CAZ 64 mg/ml AZT 32 mg/ml PIPC 128 mg/ml IPM 0.12 mg/ml 1 K. pneumoniae K. pneumoniae CLSI cefotaxime CAZ 12 Vol. 21 No

3 TEM-132 ESBL Klebsiella pneumoniae K. pneumoiae CTX-M- TEM- SHV- b- 3. PCR ESBL 25 PCR ESBL TEM- bla TEM PFGE PFGE 25 1 P1 P DNA DNA (P3) P P TEM (71.4 ), 2 4 (19.1 ), 6 2 (9.5 ) (56 ) 9 (36 ) (42.9 ), (47.6 ), CTM 4 CAZ 8 CTM, CAZ TEM-132 Vol. 21 No

4 14 2 PFGE 1 5 A P1 2 5 B P2 3 5 C P3 4 5 D P3 5 5 E P3 6 5 F P3 7 5 G P3 8 5 H P3 9 5 H P I P J P K P L P K P M P N P O P P P Q P R P S P S P M P T P U P ESBL III. K. pneumoniae K. pneumoniae ESBL 14 Vol. 21 No

5 TEM-132 ESBL Klebsiella pneumoniae 15 SHV-12 SHV-24 TEM-91 ESBL K. pneumoniae 9), 10) 25 TEM-132 TEM-132 ESBL TEM-1 b- 3 (R164H, I173V, E240 K) K. pneumoniae 11) PFGE ESBL 2 6 ESBL 12) CAZ- K. pneumoniae CAZ- K. pneumoniae ICT ESBL ICT ) de Oliveira Garcia, D., Y. Doi, D. Szabo, J. M. Adams-Haduch, T. M. Vaz, D. Leite, M. C. Padoveze, M. P. Freire, F. P. Silveira, D. L. Paterson Multiclonal outbreak of Klebsiella pneumoniae producing extended-spectrum b- lactamase CTX-M-2 and novel variant CTX-M- 59 in a neonatal intensive care unit in Brazil. Antimicrob. Agents Chemother. 52: ) Fang, H., C. Lundberg, B. Olsson-Liljequist, G. Hedin, E. Lindbäck, A. Rosenberg, J. Struwe Molecular epidemiological analysis of Escherichia coli isolates producing extendedspectrum b-lactamases for identification of nosocomial outbreaks in Stockholm, Sweden. J. Clin. Microbiol. 42: ) Clinical and Laboratory Standards Institute Performance standards for antimicrobial susceptibility testing. Eighteenth information supplement (M100-S18). Clinical and Laboratory Standards Institute, Wayne, PA. 4) Knothe, H., P. Shah, V. Krcmery, M. Antal, S. Mitsuhashi Transferable resistance to cefotaxime, cefoxitin, cefamandole and cefuroxime in clinicalisolates of Klebsiella pneumoniae and Serratia marcesens. Infection 11: ) Green, M., K. Barbadora Recovery of ceftazidime-resistant Klebsiella pneumoniae from pediatric liver and intestinal transplant recipients. Pediatr. Transplant. 2: ) Hibbert-Rogers, L. C., J. Heritage, D. M. Gascoyne-binzi, P. M. Hawkey, N. Todd, I. J. Lewis, C. Bailey Molecular epidemiology of ceftazidime resistant Enterobacteriaceae from patients on a paediatric oncology ward. J. Antimicrob. Chemother. 36: ) Shibata, N., H. Kurokawa, Y. Doi, T. Yagi, K. Yamane, J. Wachino, S. Satowa, K. Kimura, S. Ishikawa, H. Kato, Y. Ozawa, K. Shibayama, K. Kai, F. Konda, Y. Arakawa PCR classification of CTX-M-type b-lactamase genes identified in clinically isolated Gram-negative bacilli Vol. 21 No

6 16 in Japan. Antimicrob. Agents Chemother. 50: ) Duck, W. M., C. D. Steward, S. N. Banerjee, J. E. McGowan, Jr., F. C. Tenover Optimization of computer software settings improves accuracy of pulsed-field gel electrophoresis macrorestriction fragment pattern analysis. J. Clin. Microbiol. 41: ) Kurokawa H., T. Yagi, N. Shibata, K. Shibayama, K. Kamachi, Y. Arakawa A new SHV-derived extended-spectrum b-lactamase (SHV-24) that hydrolyzes ceftazidime through a single-amino-acid substitution (D179G) in the W-loop. Antimicrob. Agents Chemother. 44: ) Kurokawa, H., N. Shibata, Y. Doi, K. Shibayama, K. Kamachi, T. Yagi, Y. Arakawa A new TEM-derived extended-spectrum b-lactamase (TEM-91) with an R164C substitution at the W-loop confers ceftazidime resistance. Antimicrob. Agents Chemother. 47: ) Zarnayova, M., E. Siebor, A. Pechinot, J. M. Duez, H. Bujdakova, R. Labia, C. Neuwirth Survey of Enterobacteriaceae producing extended-spectrum b-lactamases in a Slovak hospital: Dominance of SHV-2a and characterization of TEM-132. Antimicrob. Agents Chemother. 49: ) Paterson, D. L., R. A. Bonomo Extendedspectrum b-lactamase: A clinical update. Clin. Microbiol. Rev. 18: TEM-132-producing Ceftatidime-resistant Klebsiella pneumoniae That Cause a Probable Healthcare Associated Transmission Maho Takahashi 1), Naohiro Shibata 2), Kimiko Ooya 1), Misako Horimizu 1), Kazuhiko Nagai 3), Yoshichika Arakawa 2) 1) Department of Clinical Laboratory, kaetsu Hospital, Niigata, Japan 2) Department of Bacterial Pathogenic and Infection Control, National Instiute of Infection Deseases 3) Department of Pharmacy, Kaetsu Hospital, Niigata, Japan Ceftazidime (CAZ)-resistant Klebsiella pneumoniae strains were isolated from 95 inpatients during April, 2003 and November, To characterize the molecular mechanisms of the CAZ-resistance, PCR detection of bla genes, nucleotide sequencing of the PCR products were performed for the 21 isolates, as well as pulsed-field electrophoresis (PFGE). All isolates were found to harbor bla TEM-132 by PCR amplification and nucleotide sequence analyses. By PFGE analysis, 19 isolates demonstrated the same PFGE profile suggesting a probable healthcare associated transmission of TEM-132-producing K. pneumoniae. To cope with this outbreak, the contact precaution, together with the standard precaution and enhanced sanitation of hospital environment, was performed, and we finally succeeded in the prevention of further transmission of the CAZ-resistant K. pneumoniae. Most ESBLs found in CAZ-resistant K. pneumoniae have been SHV-derived ESBLs, but the enzyme identified in the CAZ-resistant K. pneumoniae was TEM-132 that has never been reported in Japan. 16 Vol. 21 No