{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T06:04:59Z","timestamp":1759989899491},"reference-count":49,"publisher":"Microbiology Society","issue":"9","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2010,9,1]]},"abstract":"<jats:p>This study was conducted to detect and analyse the presence of extended-spectrum <jats:italic>\u03b2<\/jats:italic>-lactamase (ESBL)-producing <jats:italic>Klebsiella pneumoniae<\/jats:italic> associated with a nosocomial outbreak at a Croatian hospital. During 2007, 162 <jats:italic>K. pneumoniae<\/jats:italic> isolates with reduced susceptibility to third-generation cephalosporins were collected from various hospital units and patient specimens. Most of the strains were isolated from urine (61\u200a%), followed by blood cultures (13\u200a%), wound swabs (13\u200a%), tracheal aspirates (5\u200a%), intra-abdominal abscess aspirates (4\u200a%), intravascular catheters (3\u200a%) and cerebrospinal fluid (1\u200a%). Medical wards were the most important source of the isolates (46\u200a%); 21\u200a% of the isolates originated from surgical intensive-care units. All patients had infections acquired during their stay in hospital. No community-acquired infections were reported. Sixty of these isolates were chosen for further analysis. A double-disc synergy test (DDST) was used to detect ESBLs. MICs were determined by the broth microdilution method according to CLSI guidelines. The transferability of ceftazidime resistance was tested by conjugation (broth mating method). PCR was used to detect alleles encoding ESBL enzymes. Plasmids encoding ESBLs were extracted with the Macherey Nagel Mini kit according to the manufacturer's recommendations. The genotypes of the strains were compared by analysis of banding patterns generated by PFGE of <jats:italic>Xba<\/jats:italic>I-digested genomic DNA. ESBLs were found by DDST in all isolates. All strains were resistant to cefuroxime, ceftazidime, cefotaxime, ceftriaxone, aztreonam, piperacillin\/tazobactam and ciprofloxacin. There was variable susceptibility\/resistance to cefepime and gentamicin. No resistance to ceftazidime\/clavulanate and carbapenems was observed. Only six strains transferred resistance to an <jats:italic>Escherichia coli<\/jats:italic> recipient strain, with low frequency. All isolates yielded an amplicon of 545\u2005bp with consensus MA primers. Multiplex PCR was positive for group 1 CTX-M <jats:italic>\u03b2<\/jats:italic>-lactamases. Sequencing of selected amplicons revealed the presence of <jats:italic>bla<\/jats:italic>\n                  <jats:sub>CTX-M-15<\/jats:sub>, with coding regions containing identical nucleotide sequences. Similarly to isolates from India, our isolates contained the IS<jats:italic>EcpI<\/jats:italic> insertion sequence located upstream of the <jats:italic>bla<\/jats:italic>\n                  <jats:sub>CTX-M-15<\/jats:sub> gene, which has recently been demonstrated to mobilize 3\u2032-adjacent genes to transfer between DNA replicons. The isolates contained a large plasmid of approximately 150\u2005kb. The isolates were assigned to five clusters (&gt;85\u200a% similarity), which contained subclusters. The results of this work provided insights into the molecular epidemiology of the spread of ESBLs in <jats:italic>K. pneumoniae<\/jats:italic> involved in an outbreak at a Croatian hospital. The hospital antibiotic policy resulted in ceftriaxone being the most heavily prescribed third-generation cephalosporin, which might be expected to select for cefotaximases such as CTX-M-15.<\/jats:p>","DOI":"10.1099\/jmm.0.019778-0","type":"journal-article","created":{"date-parts":[[2010,6,25]],"date-time":"2010-06-25T03:45:50Z","timestamp":1277437550000},"page":"1069-1078","source":"Crossref","is-referenced-by-count":15,"title":["Clonal spread of CTX-M-15-producing Klebsiella pneumoniae in a Croatian hospital"],"prefix":"10.1099","volume":"59","author":[{"given":"Mirna","family":"Vranic-Ladavac","sequence":"first","affiliation":[{"name":"County of Istria Public Health Institute, Nazorova 23, Pula, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zrinka","family":"Bosnjak","sequence":"additional","affiliation":[{"name":"Clinical Hospital Center Zagreb, Kispatic Street 12, Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Natasa","family":"Beader","sequence":"additional","affiliation":[{"name":"School of Medicine, University of Zagreb, Salata 3, Zagreb, Croatia"},{"name":"Clinical Hospital Center Zagreb, Kispatic Street 12, Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nada","family":"Barisic","sequence":"additional","affiliation":[{"name":"County of Istria Public Health Institute, Nazorova 23, Pula, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Smilja","family":"Kalenic","sequence":"additional","affiliation":[{"name":"School of Medicine, University of Zagreb, Salata 3, Zagreb, Croatia"},{"name":"Clinical Hospital Center Zagreb, Kispatic Street 12, Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Branka","family":"Bedenic","sequence":"additional","affiliation":[{"name":"School of Medicine, University of Zagreb, Salata 3, Zagreb, Croatia"},{"name":"Clinical Hospital Center Zagreb, Kispatic Street 12, Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"R1","first-page":"203","article-title":"Molecular characterization by PCR-restriction fragment polymorphism of TEM beta-lactamases","volume":"134","author":"Arlet","year":"1995","journal-title":"FEMS Microbiol Lett"},{"key":"R2","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1128\/AAC.46.1.151-159.2002","article-title":"Countrywide spread of CTX-M-3 extended-spectrum beta-lactamase-producing microorganisms of the family Enterobacteriaceae in Poland","volume":"46","author":"Baraniak","year":"2002","journal-title":"Antimicrob Agents Chemother"},{"key":"R3","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1179\/joc.1998.10.6.449","article-title":"Extended-spectrum beta-lactamases in clinical isolates of Klebsiella pneumoniae from Zagreb, Croatia","volume":"10","author":"Bedenic","year":"1998","journal-title":"J Chemother"},{"key":"R4","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1007\/PL00011293","article-title":"Molecular epidemiology of extended-spectrum beta-lactamases from Klebsiella pneumoniae strains isolated in Zagreb, Croatia","volume":"20","author":"Bedenic","year":"2001","journal-title":"Eur J Clin Microbiol Infect Dis"},{"key":"R5","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1179\/joc.2005.17.4.367","article-title":"Epidemic and endemic spread of Klebsiella pneumoniae producing SHV-5 beta-lactamase in Dubrava University Hospital, Zagreb","volume":"17","author":"Bedenic","year":"2005","journal-title":"J Chemother"},{"key":"R6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/AAC.48.1.1-14.2004","article-title":"Growing group of extended-spectrum beta-lactamases: the CTX-M enzymes","volume":"48","author":"Bonnet","year":"2004","journal-title":"Antimicrob Agents Chemother"},{"key":"R7","doi-asserted-by":"crossref","first-page":"933","DOI":"10.1128\/CMR.14.4.933-951.2001","article-title":"Extended-spectrum beta-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat","volume":"14","author":"Bradford","year":"2001","journal-title":"Clin Microbiol Rev"},{"key":"R8","doi-asserted-by":"crossref","first-page":"1980","DOI":"10.1128\/AAC.42.8.1980","article-title":"CTX-M-5, a novel cefotaxime-hydrolyzing beta-lactamase from an outbreak of Salmonella typhimurim in Latvia","volume":"42","author":"Bradford","year":"1998","journal-title":"Antimicrob Agents Chemother"},{"key":"R9","doi-asserted-by":"crossref","first-page":"1237","DOI":"10.1128\/JCM.40.4.1237-1243.2002","article-title":"Epidemiology of extended-spectrum \u03b2 -lactamase-producing Enterobacter isolates in a Spanish hospital during a 12-year period","volume":"40","author":"Cant\u00f3n","year":"2002","journal-title":"J Clin Microbiol"},{"key":"R10","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1128\/AAC.46.3.630-637.2002","article-title":"Three cefotaximases, CTX-M-9, CTX-M-13, and CTX-M-14 among Enterobacteriaceae in the People's Republic of China","volume":"46","author":"Chanawong","year":"2002","journal-title":"Antimicrob Agents Chemother"},{"key":"R11","article-title":"Performance standards for antimicrobial susceptibility testing , 17th informational supplement. CLSI\/NCCLS document M100-S17","year":"2007"},{"key":"R12","doi-asserted-by":"crossref","first-page":"978","DOI":"10.1093\/jac\/dkn287","article-title":"Expansion and countrywide dissemination of ST11, ST15, and ST147 ciprofloxacin-resistant CTX-M-15-type \u03b2 -lactamase-producing Klebsiella pneumoniae epidemic clones in Hungary in 2005 \u2013 the new \u2018MRSAs'?","volume":"62","author":"Damjanova","year":"2008","journal-title":"J Antimicrob Chemother"},{"key":"R13","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1128\/AAC.48.4.1249-1255.2004","article-title":"Dissemination of CTX-M-type beta-lactamases among clinical isolates of Enterobacteriaceae in Paris, France","volume":"48","author":"Eckert","year":"2004","journal-title":"Antimicrob Agents Chemother"},{"key":"R14","doi-asserted-by":"crossref","first-page":"3724","DOI":"10.1128\/AAC.47.12.3724-3732.2003","article-title":"Prevalence and molecular epidemiology of CTX-M extended spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in Russian hospitals","volume":"47","author":"Edelstein","year":"2003","journal-title":"Antimicrob Agents Chemother"},{"key":"R15","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1128\/AAC.50.2.785-787.2006","article-title":"Emergence of Enterobacteriaceae isolates producing CTX-M extended-spectrum beta-lactamase in Austria","volume":"50","author":"Eisner","year":"2006","journal-title":"Antimicrob Agents Chemother"},{"key":"R16","first-page":"683","article-title":"The characterization of R plasmids and the detection of plasmid-specified genes","volume-title":"In Antibiotics in Laboratory Medicine","author":"Elwell","year":"1986"},{"key":"R17","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1056\/NEJMra041359","article-title":"The new beta-lactamases","volume":"352","author":"Jacoby","year":"2005","journal-title":"N Engl J Med"},{"key":"R18","doi-asserted-by":"crossref","first-page":"867","DOI":"10.1093\/clinids\/10.4.867","article-title":"Extended broad-spectrum beta-lactamases conferring transferable resistance to newer beta-lactam agents in Enterobacteriaceae : hospital prevalence and susceptibility patterns","volume":"10","author":"Jarlier","year":"1988","journal-title":"Rev Infect Dis"},{"key":"R19","first-page":"237","article-title":"Plasmid-mediated extended-spectrum beta-lactamase (CTX-M-3 like) from India and gene association with insertion sequence ISEcp1","volume":"201","author":"Karim","year":"2001","journal-title":"FEMS Microbiol Lett"},{"key":"R20","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1385\/0-89603-498-4:33","article-title":"Pulsed-field gel electrophoresis","volume-title":"In Molecular Bacteriology Protocols and Clinical Applications","author":"Kaufman","year":"1998"},{"key":"R21","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1016\/j.diagmicrobio.2007.02.005","article-title":"The emergence of a novel ceftazidime-resistant CTX-M extended-spectrum beta-lactamase, CTX-M-15, in both community-onset and hospital-acquired infections in Thailand","volume":"58","author":"Kiratisin","year":"2007","journal-title":"Diagn Microbiol Infect Dis"},{"key":"R22","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1128\/AAC.28.2.302","article-title":"Evolution of plasmid-coded resistance to broad-spectrum cephalosporins","volume":"28","author":"Kliebe","year":"1985","journal-title":"Antimicrob Agents Chemother"},{"key":"R23","doi-asserted-by":"crossref","first-page":"2855","DOI":"10.1128\/AAC.01614-06","article-title":"Extended-spectrum beta-lactamases of the CTX-M type now in Switzerland","volume":"51","author":"Lartigue","year":"2007","journal-title":"Antimicrob Agents Chemother"},{"key":"R24","doi-asserted-by":"crossref","first-page":"4224","DOI":"10.1128\/AAC.00904-06","article-title":"qnrA in CTX-M producing Escherichia coli isolates from France","volume":"50","author":"Lavigne","year":"2006","journal-title":"Antimicrob Agents Chemother"},{"key":"R25","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1093\/jac\/dkm225","article-title":"Control of extended-spectrum \u03b2 -lactamase-producing Escherichia coli and Klebsiella pneumoniae in a children's hospital by changing antimicrobial agent usage policy","volume":"60","author":"Lee","year":"2007","journal-title":"J Antimicrob Chemother"},{"key":"R26","doi-asserted-by":"crossref","first-page":"1630","DOI":"10.1128\/AAC.01431-08","article-title":"bla CTX-M genes in Escherichia coli from Croatian hospitals are located in new ( bla CTX-M-3a) and widely spread ( bla CTX-M-3a and bla CTX-M-15) genetic structures","volume":"53","author":"Literacka","year":"2009","journal-title":"Antimicrob Agents Chemother"},{"key":"R27","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1093\/jac\/dkl483","article-title":"CTX-M: changing the face of ESBLs in Europe","volume":"59","author":"Livermore","year":"2007","journal-title":"J Antimicrob Chemother"},{"key":"R28","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1111\/j.1469-0691.2004.00929.x","article-title":"Extended-spectrum beta-lactamase (ESBL) CTX-M-15-producing Escherichia coli and Klebsiella pneumoniae in Sofia, Bulgaria","volume":"10","author":"Markovska","year":"2004","journal-title":"Clin Microbiol Infect"},{"key":"R29","doi-asserted-by":"crossref","first-page":"2831","DOI":"10.1128\/JCM.00418-06","article-title":"Characterization of a large outbreak by CTX-M-1-producing Klebsiella pneumoniae and mechanisms leading to in vivo carbapenem resistance development","volume":"44","author":"Mena","year":"2006","journal-title":"J Clin Microbiol"},{"key":"R30","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1128\/AAC.00746-08","article-title":"Nationwide survey of CTX-M-type extended-spectrum \u03b2 -lactamases among Klebsiella pneumoniae isolates in Slovenian hospitals","volume":"53","author":"Me\u0161ko Megli\u010d","year":"2009","journal-title":"Antimicrob Agents Chemother"},{"key":"R31","doi-asserted-by":"crossref","first-page":"2700","DOI":"10.1128\/AAC.00068-06","article-title":"CTX-M-type extended-spectrum beta-lactamases in Italy: molecular epidemiology of an emerging countrywide problem","volume":"50","author":"Mugnaioli","year":"2006","journal-title":"Antimicrob Agents Chemother"},{"key":"R33","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1007\/BF01690097","article-title":"Detection of genes coding for extended-spectrum SHV beta-lactamases in clinical isolates by a molecular genetic method, and comparison with the E test","volume":"15","author":"N\u00fcesch-Inderbinen","year":"1996","journal-title":"Eur J Clin Microbiol Infect Dis"},{"key":"R34","doi-asserted-by":"crossref","first-page":"2359","DOI":"10.1128\/JCM.00447-06","article-title":"Spread of Escherichia coli strains with high-level cefotaxime and ceftazidime resistance between the community, long-term facilities, and hospital institutions","volume":"44","author":"Oteo","year":"2006","journal-title":"J Clin Microbiol"},{"key":"R35","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1093\/jac\/dkp211","article-title":"Emergence of CTX-M-15-producing Klebsiella pneumoniae of multilocus sequence types 1, 11, 14, 17, 20, 35 and 36 as pathogens and colonizers in newborns and adults","volume":"64","author":"Oteo","year":"2009","journal-title":"J Antimicrob Chemother"},{"key":"R36","doi-asserted-by":"crossref","first-page":"2523","DOI":"10.1128\/JCM.42.6.2523-2529.2004","article-title":"Multifocal detection of multidrug-resistant Pseudomonas aeruginosa producing PER-1 extended-spectrum beta-lactamase in Northern Italy","volume":"42","author":"Pagani","year":"2004","journal-title":"J Clin Microbiol"},{"key":"R37","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1093\/jac\/44.4.489","article-title":"Concurrent outbreaks of extended-spectrum beta-lactamase-producing organisms of the family Enterobacteriaceae in a Warsaw hospital","volume":"44","author":"Palucha","year":"1999","journal-title":"J Antimicrob Chemother"},{"key":"R38","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1093\/jac\/dki166","article-title":"Emergence of Enterobacteriaceae producing extended-spectrum beta-lactamases in the community","volume":"56","author":"Pitout","year":"2005","journal-title":"J Antimicrob Chemother"},{"key":"R39","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1093\/jac\/dkh323","article-title":"CTX-M enzymes are the most common extended-spectrum beta-lactamases among Escherichia coli in a tertiary Greek hospital","volume":"54","author":"Pournaras","year":"2004","journal-title":"J Antimicrob Chemother"},{"key":"R40","doi-asserted-by":"crossref","first-page":"2864","DOI":"10.1128\/AAC.47.9.2864-2867.2003","article-title":"Extended-spectrum beta-lactamases in Buenos Aires, Argentina, public hospitals","volume":"47","author":"Quinteros","year":"2003","journal-title":"Antimicrob Agents Chemother"},{"key":"R41","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1016\/S1473-3099(06)70599-0","article-title":"The worldwide emergence of plasmid-mediated quinolone resistance","volume":"6","author":"Robicsek","year":"2006a","journal-title":"Lancet Infect Dis"},{"key":"R42","doi-asserted-by":"crossref","first-page":"2872","DOI":"10.1128\/AAC.01647-05","article-title":"qnr prevalence in ceftazidime-resistant Enterobacteriaceae isolates from the United States","volume":"50","author":"Robicsek","year":"2006b","journal-title":"Antimicrob Agents Chemother"},{"key":"R43","first-page":"161","article-title":"Diversity of CTX-M beta-lactamases and their promoter regions from Enterobacteriaceae isolated in three Parisian hospitals","volume":"209","author":"Saladin","year":"2002","journal-title":"FEMS Microbiol Lett"},{"key":"R44","doi-asserted-by":"crossref","first-page":"2233","DOI":"10.1128\/JCM.33.9.2233-2239.1995","article-title":"Interprinting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing","volume":"33","author":"Tenover","year":"1995","journal-title":"J Clin Microbiol"},{"key":"R45","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1179\/joc.2007.19.1.97","article-title":"First report of CTX-M producing isolates from Croatia","volume":"19","author":"Tonkic","year":"2007","journal-title":"J Chemother"},{"key":"R46","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1016\/j.ijantimicag.2004.09.007","article-title":"First isolation of CTX-M-3 beta-lactamase producer in the United Kingdom","volume":"24","author":"Winstanley","year":"2004","journal-title":"Int J Antimicrob Agents"},{"key":"R47","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1093\/jac\/dkh424","article-title":"Community and hospital spread of Escherichia coli producing CTX-M extended-spectrum beta-lactamases in the UK","volume":"54","author":"Woodford","year":"2004","journal-title":"J Antimicrob Chemother"},{"key":"R48","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1016\/j.ijantimicag.2006.01.004","article-title":"Multiplex PCR for genes encoding prevalent OXA carbapenemases","volume":"27","author":"Woodford","year":"2006","journal-title":"Int J Antimicrob Agents"},{"key":"R49","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1093\/jac\/dkg103","article-title":"Production of CTX-M-3 extended-spectrum beta-lactamase and IMP-1 metallo- \u03b2 -lactamase by five Gram-negative bacilli: survey of clinical isolates from seven laboratories collected in 1998 and 2000, in the Kinki region of Japan","volume":"51","author":"Yamasaki","year":"2003","journal-title":"J Antimicrob Chemother"},{"key":"R50","doi-asserted-by":"crossref","first-page":"1098","DOI":"10.1128\/AAC.46.4.1098-1100.2002","article-title":"First description of Klebsiella pneumoniae harboring CTX-M- \u03b2 -lactamases (CTX-M-14 and CTX-M-3) in Taiwan","volume":"46","author":"Yu","year":"2002","journal-title":"Antimicrob Agents Chemother"}],"container-title":["Journal of Medical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.019778-0?crawler=true","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,9,30]],"date-time":"2020-09-30T14:40:27Z","timestamp":1601476827000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.019778-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,9,1]]},"references-count":49,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2010,9,1]]}},"URL":"https:\/\/doi.org\/10.1099\/jmm.0.019778-0","relation":{},"ISSN":["0022-2615","1473-5644"],"issn-type":[{"value":"0022-2615","type":"print"},{"value":"1473-5644","type":"electronic"}],"subject":[],"published":{"date-parts":[[2010,9,1]]}}}