Klebsiella pneumoniae carbapenemase (KPC) in urinary infection isolates

Bedenić, Branka; Sardelić, Sanda; Bogdanić, Maja; Zarfel, Gernot; Beader, Nataša; Šuto, Sandra; Krilanović, Marija; Vraneš, Jasmina

doi:10.1007/s00203-020-02161-x

Original Paper
Published: 28 January 2021

Klebsiella pneumoniae carbapenemase (KPC) in urinary infection isolates

Branka Bedenić ORCID: orcid.org/0000-0002-5946-7997^1,2,
Sanda Sardelić³,
Maja Bogdanić⁴,
Gernot Zarfel⁵,
Nataša Beader^1,2,
Sandra Šuto⁶,
Marija Krilanović⁷ &
Jasmina Vraneš^1,6

Archives of Microbiology volume 203, pages 1825–1831 (2021)Cite this article

284 Accesses
Metrics details

Abstract

Recently, emergence of carbapenem-resistance, in particular due to Klebsiella pneumoniae carbapenemase (KPC), was observed among K. pneumoniae causing urinary tract infections in Croatia. The aim of the study was to characterize, antimicrobial susceptibility, carbapenem resistance, virulence traits and plasmid types of the urinary KPC positive isolates of K. pneumoniae. The antimicrobial susceptibility to a wide range of antibiotics was determined by broth microdilution method. The transferability of meropenem resistance was determined by conjugation (broth mating method) employing Escherichia coli J63 strain resistant to sodium azide. Genes encoding broad and extended-spectrum β-lactamases, plasmid-mediated AmpC β-lactamases, group A and B carbapenemases, and carbapenem hydrolyzing oxacillinases (bla_OXA-48like), respectively, were determined by Polymerase chain reaction (PCR). In total 30 KPC-positive K. pneumoniae urinary isolates collected from different regions of Croatia were analysed. The isolates were uniformly resistant to all tested antibiotics except for variable susceptibility to gentamicin, sulphamethoxazole/trimethoprim, and colistin, respectively. Four isolates were resistant to colistin with MICs values ranging from 4 to 16 mg/L. All tested isolates were susceptible to ceftazidime/avibactam. Sixteen isolates transferred meropenem resistance to E. coli recipient strain by conjugation. Other resistance markers were not co-transferred. PCR was positive for bla_KPC and bla_SHV genes in all isolates whereas 13 isolates tested positive also for bla_TEM genes. PCR based replicon typing (PBRT) revealed the presence of FIIs in 13 and FIA plasmid in two strains. The study showed dissemination of KPC-producing K. pneumoniae in urinary isolates, posing a new epidemiological and treatment challenge. Sulphamethoxazole/trimethoprim, colistin, and ceftazidime/avibactam remain so far, as the therapeutic options.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

Arlet G, Brami G, Decre D, Flippo A, Gaillot O, Lagrange PH et al (1995) Molecular characterization by PCR restriction fragment polymorphism of TEM β-lactamases. FEMS Microbiol Lett 134:203–208. https://doi.org/10.1111/j.1574-6968.1995.tb07938.x
CAS Article PubMed Google Scholar
Avgoulea K, Di Pilato V, Zarkotou O, Sennati S, Politi L, Cannatelli A et al (2018) Characterization of extensively drug-resistant or pandrug resistant sequence type 147 and 101 OXA-48-producing Klebsiella pneumoniae causing bloodstream infections in patients in intensive care unit. Antimicrob Agents Chemother. https://doi.org/10.1128/AAC.02457-17
Article PubMed PubMed Central Google Scholar
A Baraniak, A Grabowska, R Izdebski, J Fiett, M Herda, K Bojarska et al. KPC-PL Study Group (2011) Molecular characteristics of KPC-producing Enterobacteriaceae at the early stage of their dissemination in Poland, 2008–2009. Antimicrob Agents Chemother 55(12):5493–5499. https://doi.org/10.1128/AAC.05118-11
CAS Article Google Scholar
Bedenić B, Mazzariol A, Plečko V, Bošnjak Z, Barl P, Vraneš J et al (2012) First report of KPC-producing Klebsiella pneumoniae in Croatia. J Chemother 24(4):237–239. https://doi.org/10.1179/1973947812Y.0000000017
Article PubMed Google Scholar
Bedenić B, Zujić Atalić V, Jajić I, Đjuras-Cuculić B, Godič-Torkar K, Vraneš J et al (2015) Clonal spread of Klebsiella pneumoniae producing KPC-2 β-lactamase in Croatian University Hospital. J Chemother 27(4):241–245. https://doi.org/10.1179/1973947814Y.0000000191
Article PubMed Google Scholar
Bedenić B, Sardelić S, Luxner J, Bošnjak Z, Varda-Brkić D, Lukić-Grlić A et al (2016) Molecular characterization of class B carbapenemases in advanced stage of dissemination and emergence of class D carbapenemases in Enterobacteriaceae from Croatia. Infect Genetic Evol 43(43):74–82. https://doi.org/10.1016/meegid.2016.05.011
Article Google Scholar
Bedenić B, Slade M, Žele-Starčević L, Sardelić S, Vranić-Ladavac M, Benčić A et al (2018) Epidemic spread of OXA-48 β-lactamase in Croatia. J Med Microbiol 67(8):1031–1034. https://doi.org/10.1099/jmm.0.000777
CAS Article PubMed Google Scholar
Bevivino A, Dalmastri C, Tabacchioni S, Chirarinni L, Belli ML, Piana S et al (2002) Burkholderia cepacia complex bacteria from clinical and environmental sources in Ital: genomorvar status and distribution of traits related to virulence and transmissibility. J Clin Microbiol 40(3):846–851
Article Google Scholar
Bonomo RA, Burd EM, Conly J, Limbago BM, Poirel L, Segre JA, Westblade LF (2018) Carbapenemase-producing organisms: a global scourge. Clin Infect Dis. https://doi.org/10.1093/cid/cix893
Article PubMed PubMed Central Google Scholar
Cannateli A, Giani T, D’Andrea MM, Di Pilato V, Arena F, Conte V et al (2014) MgrB inactivation is a common mechanism of colistin resistance in KPC-producing Klebsiella pneumoniae of clinical origin. Antimicrob Agents Chemother 58(10):5696–5703. https://doi.org/10.1128/AAC.03110-14
CAS Article Google Scholar
Canton R, Akova M, Carmeli Y, Giske CG, Glupczynski Y et al (2012) The European Network on carbapenemases, Rapid evolution and spread of carbapenemases among Enterobacteriaceae in Europe. Clin Microbiol Infect 18:413–431
CAS Article Google Scholar
Carattoli A, Bertini A, Villa L, Falbo V, Hopkins KL, Threlfall EJ (2005) Identification of plasmids by PCR-based replicon typing. J Microbiol Methods 63(3):219–228. https://doi.org/10.1016/j.mimet.2005.03.018
CAS Article PubMed Google Scholar
Carattoli A, Sieffert S, Schwendener S, Perreten V, Endimiani A (2015) Differentiation of IncL and IncM plasmids associated with the spread of clinically relevant antimicrobial resistance. PLoS ONE. https://doi.org/10.1371/journal.pone.0123063
Article PubMed PubMed Central Google Scholar
Clinical and Laboratory Standards Institute (2018) Performance Standards for Antimicrobial Susceptibility Testing. 28th ed. CLSI supplement. M100. Wayne, PA: CLSI
Coudron PE (2005) Inhibitor based methods for detection of plasmid-mediated AmpC β-Lactamase in Klebsiella spp Escherichia coli, and Proteus mirabilis. J Clin Microbiol 43(8):4163–4167. https://doi.org/10.1128/JCM.43.8.4163-4167.2005
CAS Article PubMed PubMed Central Google Scholar
Elwell LP, Falkow S (1986) The characterization of R plasmids and the detection of plasmid-specified genes. In: Lorian V (ed) Antibiotics in laboratory medicine, 2nd edn. Williams and Wilkins, Baltimore MD, pp 683–721
Google Scholar
European Committee for Antimicrobial Susceptibility Testing (2019) Breakpoint Tables for interpreting MICs and zone diamaters. Version 9.0. Available at https://www.eucast.org
Giani T, Pini B, Arena F, Conte V, Bracco S, Migliavacca R, Pantosti A, Luzzaro F, Rossolini GM, The AMLI-CRE Survey Participants (2013) Epidemic diffusion of KPC-carbapenemase-producing Klebsiella pneumoniae in Italy: results of the first countrywide survey, 15 May to 30 June 2011. Euro Surveillance. https://doi.org/10.2807/ese.18.22.20489-en
Article PubMed Google Scholar
Huang TD, Bogaerts P, Berhin M, Hoebeke M, Bauraing C, Glupczyinski Y et al (2015) Increasing proportion of carbapenemase-producing Enterobacteriaceae and emergence of MCR-I producer through a multicenter study among hospital based and private laboratories in Belgium from September to November 2015. Euro Surveill. https://doi.org/10.2807/1560-7917.ES.2017.22.19.30530
Article Google Scholar
Jarlier V, Nicolas MH, Fournier G, Philippon A (1988) Extended broad-spectrum β-lactamases conferring transferable resistance to newer β-lactam agents in Enterobacteriaceae: hospital prevalence and susceptibility patterns. Rev Infect Dis 10(4):867–878
CAS Article Google Scholar
Jelić M, Butić I, Plečko V, Cipriš I, Jajić I, Bejuk D et al (2016) (2016) KPC-producing Klebsiella pneumoniae isolates in Croatia: a nationwide survey. Microb Drug Resist 22(8):662–667. https://doi.org/10.1089/mdr.2015.0150
CAS Article PubMed Google Scholar
Kim SK, Hong SG, Moland ES, Thomson KS (2007) Convenient test using combination of chelating agents for detection of metallo-β-lactamases in clinical laboratory. J Clin Microbiol 45(9):2798–2801. https://doi.org/10.1128/JCM.02486-06
CAS Article PubMed PubMed Central Google Scholar
Krilanović M, Tomić-Paradžik M, Meštrović T, Beader N, Herljević Z, Conzemius R et al (2020) Extended-spectrum beta-lactamases and plasmid diversity in urinary isolates of Escherichia coli in Croatia: a nation-wide, multicentric, retrospective study. Folia Microbiol. https://doi.org/10.1007/s12223-019-00769-1
Article Google Scholar
Liu Liu YY, Wang Y, Walsh T, Yi LX, Zhang R, Spencer J et al (2016) Emergence of plasmid-mediated colistin resistance mechanism MCR-I in animals and human beings in China: a microbiological and molecular biological study. Lancet Infect Dis 6(2):161–168. https://doi.org/10.1016/S1473-3099(15)00424-7
CAS Article Google Scholar
Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG et al (2012) (2012) Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 18(3):268–281. https://doi.org/10.1111/j.1469-0691.2011.03570.x
CAS Article PubMed Google Scholar
Mammina C, Bonura C, Bernardo FD, Aleo A, Fascina T, Sodano C et al (2012) Ongoing spread of colistin-resistant Klebsiella pneumoniae in different wards on an acute general hospital, Italy, June to December 2011. Euro Surveillance. https://doi.org/10.2807/ese.17.33.20248-en
Article PubMed Google Scholar
Nordmann P, Poirel L (2019) Epidemiology and diagnostics of carbapenem resistance in gram-negative bacteria. Clin Infect Dis. https://doi.org/10.1093/cid/ciz824
Article PubMed PubMed Central Google Scholar
Nüesch-Inderbinen MT, Hächler H, Kayser FH (1996) Detection of genes coding for extended-spectrum SHV β-lactamases in clinical isolates by a molecular genetic method, and comparison with the E test. Eur J Clin Microbiol Infect Dis 15(5):398–402
Article Google Scholar
Pagani L, Mantengoli E, Migliavacca R, Nucleo E, Pollini S, Spalla M et al (2004) Multifocal detection of multidrug-resistant Pseudomonas aeruginosa producing PER-I extended-spectrum β-lactamase in Northern Italy. J Clin Microbiol 42(6):2523–2529. https://doi.org/10.1128/JCM.42.6.2523-2529.2004
CAS Article PubMed PubMed Central Google Scholar
Pasteran F, Mendez T, Guerriero L, Rapoport M, Corso A (2009) Sensitive screening test for suspected class a carbapenemase production in species of Enterobacteriaceae. J Clin Microbiol 47(6):1631–1639. https://doi.org/10.1128/JCM.00130-09
CAS Article PubMed PubMed Central Google Scholar
Perez-Perez FJ, Hanson ND (2002) Detection of plasmid-mediated AmpC β-lactamase genes in clinical isolates by using multiplex PCR. J Clin Microbiol 40(6):2153–2162. https://doi.org/10.1128/JCM.40.6.2153-2162.2002
CAS Article PubMed PubMed Central Google Scholar
Petrosillo N, Vranić-Ladavac M, Feudi C, Villa L, Fortini D, Barišić N et al (2015) Spread of Enterobacter cloacae carrying bla_NDM-1, bla_CTX-M-15, bla_SHV-12 and plasmid-mediated quinolone resistance genes in a surgical intensive care unit in Croatia. J Glob Antimicrob Res 4:44–48. https://doi.org/10.1016/j.jgar.2015.09.008
Article Google Scholar
Poirel L, Walsh TR, Cuveiller V, Nordman P (2011) Multiplex PCR for detection of acquired carbapenemases genes. Diagn Microbiol Infect Dis 70(1):119–123. https://doi.org/10.1016/j.diagmicrobio.2010.12.002
CAS Article PubMed Google Scholar
Robicsek A, Jacoby GA, Hooper DC (2006) The worldwide emergence of plasmid-mediated quinolone resistance. Lancet Infect Dis 6(10):629–640. https://doi.org/10.1016/S1473-3099(06)70599-0
CAS Article PubMed Google Scholar
Saladin M, Cao VT, Lambert T, Donay JL, Herrmann JL, Ould-Hocine Z et al (2002) Diversity of CTX-M β-lactamases and their promoterregions from Enterobacteriaceae isolated in three Parisian hospitals. FEMS Microbiol Lett. https://doi.org/10.1111/j.1574-6968.2002.tb11126.x
Article PubMed Google Scholar
Schiller NL, Hatch RA (1983) The serum sensitivity, colony morphology, serogroup specificity, and outer membrane protein of Pseudomonas aeruginosa strains isolated from several clinical sites. Diagn Microbiol Infect Dis 1:145–147
CAS Article Google Scholar
Van Duin D, Doi Y (2017) The global epidemiology of carbapenemase-producing Enterobacteriaceae. Virulence 8(4):460–469. https://doi.org/10.1080/21505594.2016.1222343
CAS Article PubMed Google Scholar
Vargas JM, Mochi MP, Nuñez JM, Cáceres M, Mochi S, Del Campo MR, Jure MA (2019) Virulence factors and clinical patterns of multiple-clone hypermucoviscous KPC-2 producing K. pneumonia. Heliyon. https://doi.org/10.1016/j.heliyon.2019.e01829
Article PubMed PubMed Central Google Scholar
Woodford N, Ward ME, Kaufmann ME, Turton J, Fagan EJ, James D et al (2004) Community and hospital spread of Escherichia coli producing CTX-M extended-spectrum β-lactamases in the UK. J Antimicrob Chemother 54(4):735–743. https://doi.org/10.1093/jac/dkh424
CAS Article PubMed Google Scholar
Woodford N, Fagan EJ, Ellington MJ (2006) Multiplex PCR for rapid detection of genes encoding CTX-M extended-spectrum β-lactamases. J Antimicrob Chemother 57(1):154–155. https://doi.org/10.1093/jac/dki412
CAS Article PubMed Google Scholar
Yao B, Xiao X, Zhout X, Zhang I (2015) Clinical and molecular characteristics of multi-clone carbapenem-resistant hypervirulent isolates in tertiary hospital in Bejing, China. Int J Infect Dis 37:107–112. https://doi.org/10.1016/j.ijid.2015.06.023
Article PubMed Google Scholar
Zujić Atalić V, Bedenić B, Kocsis E, Mazzariol A, Sardelić S, Barišić M et al (2014) Diversity of carbapenemases in clinical isolates of Enterobacteriaceae in Croatia-the results of the multicenter study. Clin Microbiol Infect 20(11):O894-903. https://doi.org/10.1111/1469-0691.12635
CAS Article PubMed Google Scholar

Download references

Author information

Affiliations

School of Medicine, University of Zagreb, Zagreb, Croatia
Branka Bedenić, Nataša Beader & Jasmina Vraneš
Clinical Department for Clinical and Molecular Microbiology, University Hospital Center Zagreb, Kišpatić Street 12, Zagreb, Croatia
Branka Bedenić & Nataša Beader
University Hospital Center Split, Split, Croatia
Sanda Sardelić
National Public Health Institute, Zagreb, Croatia
Maja Bogdanić
Institute for Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, Austria
Gernot Zarfel
Andrija Štampar Teaching Public Health Institute, Zagreb, Croatia
Sandra Šuto & Jasmina Vraneš
Public Health Institute of Dubrovnik-Neretva County, Dubrovnik, Croatia
Marija Krilanović

Authors

Branka Bedenić
View author publications
You can also search for this author in PubMed Google Scholar
Sanda Sardelić
View author publications
You can also search for this author in PubMed Google Scholar
Maja Bogdanić
View author publications
You can also search for this author in PubMed Google Scholar
Gernot Zarfel
View author publications
You can also search for this author in PubMed Google Scholar
Nataša Beader
View author publications
You can also search for this author in PubMed Google Scholar
Sandra Šuto
View author publications
You can also search for this author in PubMed Google Scholar
Marija Krilanović
View author publications
You can also search for this author in PubMed Google Scholar
Jasmina Vraneš
View author publications
You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Branka Bedenić.

Ethics declarations

Conflict of interest

The authors do not declare any conflict of interest.

Ethical statement

The Ethical permission was not necessary. The study did not involve human or animal subjects. It is in vitro study. Urine samples were collected for purpose of routine diagnostic and informed consent was not necessary.

The part of the results is in shown in European Congress for Clinical Microbiology and Infectious Diseases (ECCMID) 2020 online library (the congress was cancelled).

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Communicated by Erko Stackebrandt.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bedenić, B., Sardelić, S., Bogdanić, M. et al. Klebsiella pneumoniae carbapenemase (KPC) in urinary infection isolates. Arch Microbiol 203, 1825–1831 (2021). https://doi.org/10.1007/s00203-020-02161-x

Download citation

Received: 16 April 2020
Revised: 25 November 2020
Accepted: 23 December 2020
Published: 28 January 2021
Issue Date: May 2021
DOI: https://doi.org/10.1007/s00203-020-02161-x

Keywords

Klebsiella pneumoniae
KPC
Urinary tract infections
Resistance