Hospital Acquired Infections by Carbapenem Resistant Enterobacteriacea

Document Type : Original Article

Authors

1 Departments of 1Medical Microbiology and Immunolog ,Faculty of Medicine, Sohag University

2 Department of anesthesia and ICU,Sohag University, sohag,Egypt

3 Departments of Medical microbiology and immunology و faculty of medicine ,sohag university

Abstract

CRE) infections have become an increasing concern for healthcare services worldwide. Community and hospital-acquired infections caused by these bacteria have been associated with significant morbidity and mortality with limited treatment options. Rapid and accurate detection of carbapenem resistance in these bacteria is important for infection control. Objectives: To detect the prevalence of carbapenem resistant Enterobacteriaceae (CRE) species and determine their antimicrobial susceptibility profile using the Vitek 2 system and detect the presence of carbapenemases genes using Multiplex PCR. Methodology: Various clinical samples were collected from 469 patients admitted in Sohag university hospitals in the period between April 2016 and August 2017, CRE isolates were identified by conventional methods and antimicrobial susceptibility testing using disc diffusion method and also performed by Vitek 2 automated system, Multiplex PCR was used for detection of carbapenemases genes as blaKPC, blaVIM, blaIMP, blaNDM-1 and blaOXA-48. Results: The prevalence of carbapenem resistant Enterobacteriaceae (CRE) species was 19.9%, Klebsiella pneumoniae was the most common species (51.4%), Escherichia Coli (28.6%), Enterobacter aerogenes(8.6%), Acinetobacter baumannii (5.7%), Vitek 2 system identified CRE isolates with 82.7% sensitivity, 98.6 % specificity and 90.6% diagnostic accuracy, 25.7 % of CRE strains were isolated from the internal ICU and 20 % from chest department and mostly isolated from urine(40%) and from endotracheal tubes swabs(28.6%), 77.1% of CRE isolates contained carbapenemases genes, 62.1 % were blaKPC positive, 20.7 % were blaVIM-positive, 3.4 % were blaNDM-positive, 13.8 % were blaOXA-48-positive and none was blaIMP-positive. Conclusion:  Conventional methods supported by Vitek 2 system is a valuable method for identification of CRE species, the detected carbapenemases genes in this study indicates that carbapenem resistance is spreading in Egypt and support the use of molecular methods for the rapid detection of CRE for successful implementation of infection control measures, we recommend routine testing to determine carbapenem resistance in Enterobacteriaceae in health facilities in Egypt.
Keywords: Carbapenem, Enterobacteriacea, Vitek 2, Multiplex PCR
Corresponding author: Tamer Mohamed Mahmoud, Department of Medical Microbiology and Immunology, Sohag University. E-mail: tamermmm2000@yahoo.com

Wang JT, Wu UI, Lauderdale TL, Chen MC, Li SY, Hsu LY, et al. Carbapenem-nonsusceptible Enterobacteriaceae in Taiwan. PLoS One. 2015;10:e0121668.
2. Nordmann P, Naas T, Poirel L. Global spread of carbapenemase-producing Enterobacteriaceae. Emerg Infect Dis. 2011;17:1791–8.
3. Demir Y, Zer Y, Karaoglan I. Investigation of VIM, IMP, NDM-1, KPC AND OXA-48 enzymes in Enterobacteriaceae strains. Pak J Pharm Sci. 2015;28:1127–33.
4. Djahmi N, Dunyach-Remy C, Pantel A, Dekhil M, Sotto A, Lavigne JP. Epidemiology of carbapenemase-producing Enterobacteriaceae and acinetobacter baumannii in Mediterranean countries. Biomed Res Int. 2014;2014:305784.
5. Del Franco M, Paone L, Novati R, Giacomazzi CG, Bagattini M, Galotto C, et al. Molecular epidemiology of carbapenem resistant Enterobacteriaceae in Valle d’Aosta region, Italy, shows the emergence of KPC-2 producing Klebsiella pneumoniae clonal complex 101 (ST101 and ST1789). BMC Microbiol. 2015;15:260.
6. Naas T, Nordmann P Analysis of a carbapenem-hydrolyzing class A β-lactamase from Enterobacter cloacae and of its LysR-type regulatory protein. Proc Natl Acad Sci U S A. 1994;91:7693–7 10.1073/pnas.91.16.7693
7. Tsakris A, Kristo I, Poulou A, Markou F, Ikonomidis A, Pournaras S. First occurrence of KPC-2-possessing Klebsiella pneumoniae in a Greek hospital and recommendation for detection with boronic acid disc tests. J Antimicrob Chemother. 2008;62:1257–60.
8. Wei ZQ, Du XX, Yu YS, Shen P, Chen YG, Li LJ. Plasmid-mediated KPC-2 in a K. pneumoniae isolate from China. Antimicrob Agents Chemother. 2007;51:763–5.
9. Queenan AM, Bush K. Carbapenemases: the versatile beta-lactamases. Clin Microbiol Rev. 2007;20:440–58.
10.Ranjbar R, Karami A, Farshad S, Giammanco GM, Mammina C. Typing methods used in the molecular epidemiology of microbial pathogens: a how-to guide. New Microbiol. 2014;37(1):1–15.
11.Sabat AJ, Budimir A, Nashev D, Sa-Leao R, van Dijl J, Laurent F, et al. Overview of molecular typing methods for outbreak detection and epidemiological surveillance. Euro Surveill. 2013;18(4):20380.
12.Gupta N, Limbago BM, Patel JB, Kallen AJ. Carbapenem-resistant Enterobacteriaceae: epidemiology and prevention. Clin Infect Dis. 2011; 53(1): 60-7
13. Lee CR, Lee JH, Park KS, Kim YB, Jeong BC, Lee SH. Global Dissemination of Carbapenemase-Producing Klebsiella pneumoniae: Epidemiology, Genetic Context, Treatment Options, and Detection Methods. Front Microbiol. 2016; 7: 895
14.Clinical and Laboratory Standards Institute. 2013 Performance Standards for antimicrobial susceptibility testing, 23rd informational supplement. CLSI document M100-S23.
15. Martha F.Mushi, Stephen E.Mshana,Can Imirzalioglu, and Freddie Bwanga. Carbapenemase Genes among Multidrug Resistant Gram Negative Clinical Isolates from a Tertiary Hospital in Mwanza, Tanzania. Biomed Res Int. 2014; 2014: 303104.
16.Kazım Sahin, Ayse Tekin, Sule Ozdas, Demet Akin, Hande Yapislar5, Aziz Ramazan Dilek1 and Emine Sonmez: Evaluation of carbapenem resistance using phenotypic and genotypic techniques in Enterobacteriaceae isolates. Ann Clin Microbiol Antimicrob  (2015) 14:44
17. Nahed A. Elraghya, Wafaa A. Zahrana, Amal F. Makleda, Hatem M. El-Sebaeyb, Ghada R. El-Hendawya, Nahla A. Melakea, Elham Awadc, Amira H. El-Khayata: Multidrug-resistant Enterobacteriaceae nosocomial uropathogens at Menoufia University Hospitals: phenotypic characterization and detection of resistance genes using real-time PCR: Menoufia Medical Journal 2016, 29:855–86
18. Shaaban M, Ghozlan H, El Maghraby M. Susceptibility of bacteria infecting urinary tract to some antibiotics and essential oils. J Appl Pharm Sci 2012; 02:90–98.
19. Abd El –Mongy M, Reyad A. Sensitivity of urinary tract bacteria to certain antibiotics, Egypt. J Exp Biol 2013; 9:213–218.
20. Mostafa S. El-Rehewy, Ehsan Abdel Saboor, Noha A. Afifi, Maggie A. Ibrahim and  Shereen S. Qayed: Detection and Characterization of Nosocomial CarbapenemResistant Gram Negative Bacilli from Assuit University Hospitals: Egyptian Journal of Medical Microbiology  Volume 25 / No.3 / July 2016    9-17
21. El-Kazzaz S, Abou El-khier N. AmpC and metallo beta-lactamases producing Gram negative bacteria in patients with hematological malignancy. Afr J Microbiol Res 2015; 9:1247–1254.
22. Wattal C, Goel N, Oberoi JK, Raveendran R, Datta S and Prasad KJ. Surveillance of multidrug resistant organisms in a tertiary care hospital in Delhi, India. J. Assoc. Physicians India. 2010; 58:32-36.    
23.Irmak Baran and Neriman Aksu: Phenotypic and genotypic characteristics of carbapenem-resistant Enterobacteriaceae in a tertiary-level reference hospital in Turkey. Ann Clin Microbiol Antimicrob  (2016) 15:20
24.Pollett S, Miller S, Hindler J, Uslan D, Carvalho M, Humphries RM. Phenotypic and molecular characteristics of carbapenem-resistant Enterobacteriaceae in a health care system in Los Angeles, California, from 2011 to 2013. J Clin Microbiol. 2014;52:4003–9.
25.Hammoudi D, Ayoub Moubareck C, Aires J, Adaime A, Barakat A, Fayad N, et al. Countrywide spread of OXA-48 carbapenemase in Lebanon: surveillance and genetic characterization of carbapenem-non-susceptible Enterobacteriaceae in 10 hospitals over a one-year period. Int J Infect Dis. 2014;29:139–44.
26. Hamzan NI, Yean CY, Rahman RA, Hasan H, Rahman ZA. Detection of blaIMP4 and blaNDM1 harboring Klebsiella pneumoniae isolates in a university hospital in Malaysia. Emerg Health Threats J. 2015;8:26011.
27.Kucukates E and Kocazeybek B. High resistance rate against 15 different antibiotics in aerobic gramnegative bacteria isolates of cardiology intensive care unit patients. Indian. J. Med. Microbial. 2002; 20: 208-210. 
28.Dizbay M, Tunccan OG, Karasahin O, Aktas F. Emergence of carbapenemresistant Klebsiella spp infections in a Turkish university hospital: epidemiology and risk factors. J Infect Dev Ctries. 2014;8:044–9.
29. Teo J, Cai Y, Tang S, Lee W, Tan TY, Tan TT, et al. Risk factors, molecular epidemiology and outcomes of ertapenem-resistant, carbapenemsusceptible Enterobacteriaceae: a case–case–control study. PLoS One. 2012;7:e34254.
30. Girgis S, Othman H, Kassem N, Abdou S. Evaluation of boronic acid disk for detection of Klebsiella pneumonia carbapenemase in Klebsiella pneumonia. Int J Curr Microbiol App Sci 2015; 4:772–783.
31.Baroud M, Dandache I, Araj GF, Wakim R, Kanj S, Kanafani Z, et al. Underlying mechanisms of carbapenem resistance in extended spectrum betalactamase producing Klebsiella pneumoniae and Escherichia coli isolates at a tertiary care centre in Lebanon: role of OXA-48 and NDM-1 carbapenemases. Int J Antimicrob Agents. 2013;41:75–9.
 
 
 
 
32.Eser Koseoglu O, Uludag Altun H, Ergin A, Boral B, Sener B, Hascelik G. Carbapenem resistance in ESBL positive Enterobacteriaceae isolates causing invasive infections. Mikrobiyol Bul. 2014;48(1):59–69.
33.Gomez-GilMR,Pano-PardoJR,Romero-GomezMP,GasiorM,Lorenzo M, Quiles I, et al. Detection of KPC-2-producing Citrobacter freundii isolates in Spain. J Antimicrob Chemother. 2010;65(12):2695–7.
34.Nevreste Celikbilek, Ozlem Unaldi, Fisun Kirca, Aysegul Gozalan, Ziya CAcikgoz, and Riza Durmaz: Molecular Characterization of Carbapenem-ResistantKlebsiella pneumoniae Species Isolated From a Tertiary Hospital, Ankara, Turkey. JMicrobiol. 2017 August 27.
35. Iraz M, Ozad Duzgun A, Sandalli C, Doymaz MZ, Akkoyunlu Y, Saral A, et al. Distribution of beta-lactamase genes among carbapenem-resistant Klebsiella pneumoniae strains isolated from patients in Turkey. Ann Lab Med. 2015;35(6):595–601.