Characterization of KPC, NDM and VIM Type Carbapenem Resistance Enterobacteriaceae from North Eastern, Nigeria
Affiliation(s)
1 Department of Medical Microbiology, Faculty of Basic Medical Sciences, College of Health Sciences, Usmanu Danfodio University, Sokoto, Nigeria. 2 Department of Medical Microbiology, College of Medical Sciences, University of Maiduguri, Maiduguri, Nigeria. 3 Department of Medical Microbiology, Faculty of Basic Medical Sciences, College of Health Sciences, Obafemi Awolowo University, Ile-Ife, Nigeria.
1 Department of Medical Microbiology, Faculty of Basic Medical Sciences, College of Health Sciences, Usmanu Danfodio University, Sokoto, Nigeria. 2 Department of Medical Microbiology, College of Medical Sciences, University of Maiduguri, Maiduguri, Nigeria. 3 Department of Medical Microbiology, Faculty of Basic Medical Sciences, College of Health Sciences, Obafemi Awolowo University, Ile-Ife, Nigeria.
ABSTRACT
Introduction and Aim: Carbapenem resistance among species of Enterobacteriaceae has emerged as a global public health problem that adds to the high cost of care, severity and high mortality of otherwise straightforward infections. Governments around the world are devoting efforts to combat this important threat. The present study was undertaken in our setting to detect and characterize carbapenem resistance among Enterobacteriaceae. Methodology: Confirmed species of Enterobacteriaceae isolated from 225 patients that were admitted in various units of University of Maiduguri Teaching Hospital (UMTH) Maiduguri were screened for carbapenem resistance with meropenem and ertapenem disc (10 μg, Oxoid, England) using clinical and laboratory standards institute (CLSI) breakpoints. Suspected carbapenemase producers were subjected to confirmation using Modified Hodge Test method. Detection of the carbapenemase genes was done by multiplex PCR using KPC, NDM-1 and VIM primers. Results: A total of 225 clinical isolates of Enterobacteriaceae comprising 73 (32.4%) of Klebsiella pneumoniae, 61 (27.1%) of Escherichia coli, 21 (9.3%) of Proteus mirabilis, 18 (8.0%) of Klebsiella oxytoca, 13 (5.8%) of Morganella morganii, 12 (5.3%) of Citrobacter freundii, 12 (5.3%) of Serratia marcescens, 7 (3.1) of Enterobacter aerogenes, 3 (1.4%) of Klebsiella ozaenae, 3 (1.4%) of Hafnia alvei and 2 (0.9%) of Citrobcter sedlakii were isolated. A total of 28 (12.4%) of the isolates screened positive as carbapenemase producers. All the 28 screened isolates were further subjected to confirmation using the Modified Hodge Test for which 23 (10.2%) were confirmed resistant. Therefore a prevalence of 10.2% for carbapenem resistance was recorded in this study. Based on multiplex polymerase chain reaction, the various percentage genotypes of the carbapenemase producers were: 11 (47.8%) for KPC, 2 (8.7%) for VIM while 5 (21.7%) isolates have co-existence of the NDM-1 and VIM genes. However, 5 (21.7%) of the isolates have none of the genes screened for in them. The occurrence of carbapenemase genes among species of Enterobacteriaceae was as follows: 6 (33.3%) for Klebsiella pneumoniae, 4 (22.2%) for Escherichia coli, 4 (22.2%) for Proteus mirabilis, 3 (16.7%) for Serratia marcescens and then 1 (5.6%) for Klebsiella oxytoca. Conclusion: Despite the low use of carbapenem agents in the study area, carbapenem resistance was documented. This calls for an ongoing surveillance and infection control practices of this and other emerging resistance threat in all health centers of Nigeria. Irrational use of antibiotics must be discouraged so as to reduce this resistance threat. Antibiotic stewardship program should be established in all tertiary health centers of Nigeria.
Introduction and Aim: Carbapenem resistance among species of Enterobacteriaceae has emerged as a global public health problem that adds to the high cost of care, severity and high mortality of otherwise straightforward infections. Governments around the world are devoting efforts to combat this important threat. The present study was undertaken in our setting to detect and characterize carbapenem resistance among Enterobacteriaceae. Methodology: Confirmed species of Enterobacteriaceae isolated from 225 patients that were admitted in various units of University of Maiduguri Teaching Hospital (UMTH) Maiduguri were screened for carbapenem resistance with meropenem and ertapenem disc (10 μg, Oxoid, England) using clinical and laboratory standards institute (CLSI) breakpoints. Suspected carbapenemase producers were subjected to confirmation using Modified Hodge Test method. Detection of the carbapenemase genes was done by multiplex PCR using KPC, NDM-1 and VIM primers. Results: A total of 225 clinical isolates of Enterobacteriaceae comprising 73 (32.4%) of Klebsiella pneumoniae, 61 (27.1%) of Escherichia coli, 21 (9.3%) of Proteus mirabilis, 18 (8.0%) of Klebsiella oxytoca, 13 (5.8%) of Morganella morganii, 12 (5.3%) of Citrobacter freundii, 12 (5.3%) of Serratia marcescens, 7 (3.1) of Enterobacter aerogenes, 3 (1.4%) of Klebsiella ozaenae, 3 (1.4%) of Hafnia alvei and 2 (0.9%) of Citrobcter sedlakii were isolated. A total of 28 (12.4%) of the isolates screened positive as carbapenemase producers. All the 28 screened isolates were further subjected to confirmation using the Modified Hodge Test for which 23 (10.2%) were confirmed resistant. Therefore a prevalence of 10.2% for carbapenem resistance was recorded in this study. Based on multiplex polymerase chain reaction, the various percentage genotypes of the carbapenemase producers were: 11 (47.8%) for KPC, 2 (8.7%) for VIM while 5 (21.7%) isolates have co-existence of the NDM-1 and VIM genes. However, 5 (21.7%) of the isolates have none of the genes screened for in them. The occurrence of carbapenemase genes among species of Enterobacteriaceae was as follows: 6 (33.3%) for Klebsiella pneumoniae, 4 (22.2%) for Escherichia coli, 4 (22.2%) for Proteus mirabilis, 3 (16.7%) for Serratia marcescens and then 1 (5.6%) for Klebsiella oxytoca. Conclusion: Despite the low use of carbapenem agents in the study area, carbapenem resistance was documented. This calls for an ongoing surveillance and infection control practices of this and other emerging resistance threat in all health centers of Nigeria. Irrational use of antibiotics must be discouraged so as to reduce this resistance threat. Antibiotic stewardship program should be established in all tertiary health centers of Nigeria.
Cite this paper
Mohammed, Y. , Zailani, S. and Onipede, A. (2015) Characterization of KPC, NDM and VIM Type Carbapenem Resistance Enterobacteriaceae from North Eastern, Nigeria. Journal of Biosciences and Medicines, 3, 100-107. doi: 10.4236/jbm.2015.311013.
Mohammed, Y. , Zailani, S. and Onipede, A. (2015) Characterization of KPC, NDM and VIM Type Carbapenem Resistance Enterobacteriaceae from North Eastern, Nigeria. Journal of Biosciences and Medicines, 3, 100-107. doi: 10.4236/jbm.2015.311013.
References
[1] Patrice, N., Laurent, P. and Laurent, D. (2012) Rapid Detection of Carbapenemase Producing Enterobacteriaceae. Emerging Infectious Diseases, 18, 1503-1507.
http://dx.doi.org/10.3201/eid1809.120355 [2] Mammeri, H., Guillon, H.E. and Nordmann, P. (2010) Phenotypic and Biochemical Comparison of the Carbapenems Hydrolyzing Activities of Five Plasmid-Borne AmpC Beta Lactamases. Antimicrobial Agents and Chemotherapy, 54, 4556-4560.
http://dx.doi.org/10.1128/AAC.01762-09 [3] Olaitan, A.O., Berrazeg, M., Fagade, O.E., Adelowo, O.O., Alli, J.A. and Rolain, J.M. (2013) Emergence of Multi Drug Resistant Acinetobacter baumanii Producing OXA-23 Carbapenemase, Nigeria. International Journal of Infectious Diseases, 17, 469-470.
http://dx.doi.org/10.1016/j.ijid.2012.12.008 [4] Centre for Disease Control (2013) Stop Infections from Lethal CRE Germs Now. CDC Vital Signs Report.
http://www.cdc.gov/hai/organisms/cre/TrackingCRE.html [5] Winn, W., Allen, S., Janda, W., Koneman, E., Procop, G., Schreckenberger, P., et al. (2006) Introduction. Koneman’s Color Atlas and Text Book of Diagnostic Microbiology. Lippincott Williams & Wilkins, Baltimore and Philadelphia, 21-25. [6] CLSI (2011) Performance Standards for Antimicrobial Susceptibility Testing: Twenty-First Informational Supplement. CLSI Document M100-S21. Clinical and Laboratory Standard Institute, Wayne. [7] Anderson, K.F., Lonsway, D.R., Rasheed, J.K., Biddle, J., Jensen, B. and McDougal, L.K. (2007) Evaluation of Methods to Identify the Klebsiella pneumoniae Carbapenemase in Enterobacteriaceae. Journal of Clinical Microbiology, 45, 2723-2725.
http://dx.doi.org/10.1128/JCM.00015-07 [8] Van-Tongeren, S.P., Degener, J.E. and Harmsen, H.J.M. (2011) Comparison of Three Rapid and Easy Bacterial DNA Extraction Methods for Use with Quantitative Real Time PCR. European Journal of Clinical Microbiology & Infectious Diseases, 30, 1053-1061.
http://dx.doi.org/10.1007/s10096-011-1191-4 [9] Nordmann, P., Poirel, L., Carrer, A., Toleman, M.A. and Walsh, T.R. (2011) How to Detect NDM-1 Producers. Journal of Clinical Microbiology, 49, 718-721.
http://dx.doi.org/10.1128/JCM.01773-10 [10] Ellington, M.J., Kistler, J., Livermore, D.M. and Woodford, N. (2007) Multiplex PCR for Rapid Detection of Genes Encoding Acquired Metallo-Beta-Lactamases. Journal of Antimicrobial Chemotherapy, 59, 321-322.
http://dx.doi.org/10.1093/jac/dkl481 [11] Pillai, D.R., Melano, R., Rawte, P., Lo, S., Tijet, N., Fuksa, M., et al. (2009) Klebsiella pneumoniae Carbapenemase, Canada. Emerging Infectious Diseases, 15, 827-829.
http://dx.doi.org/10.3201/eid1505.081536 [12] Anna, K.M., Inneke, V.H., Flavia, R., Anna, S.L. and Silvia, F.C. (2009) Multiplex PCR for Rapid Detection of Genes Encoding Oxacillinase and Metallo-Beta-Lactamases in Carbapenem Resistant Acinotobacter Species. JMM Correspondence, 10, 1522-1524.
http://jmm.sgmjournals.org [13] Yusuf, I., Arzai, A.H., Getso, M.I., Sherif, A. and Haruna, M. (2012) Emergence of Carbapenem Resistant Enterobacteriaceae in Surgical and Intensive Care Units of a Hospital with Low Usage of Carbapenem in Kano, North West Nigeria. Antimicrobial Resistance and Infection Control, 2, 75-78.
http://dx.doi.org/10.1186/2047-2994-2-S1-P75 [14] Yusuf, I., Magashi, A.M., Firdausi, F.S., Sharif, A.A., Getso, M.I., Bala, J.A., et al. (2012) Phenotypic Detection of Carbapenemases in Members of Enterobacteriacea in Kano, Nigeria. International Journal of Science and Technology, 2, 802-806. [15] Jones, R.N., Biedenbach, D.J., Sader, H.S., Fritsche, T.R., Toleman, M.A. and Walsh, T.R. (2005) Emerging Epidemic of Metallo-Beta Lactamases Mediated Resistance. Diagnostic Microbiology and Infectious Diseases, 51, 77-84.
http://dx.doi.org/10.1016/j.diagmicrobio.2004.07.003 [16] Landman, D., Bratu, S., Kochar, S., Panwar, M., Trehan, M., Doymaz, M. and Quale, J. (2007) Evolution of Antimicrobial Resistance among Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae in Brooklyn, NY. Journal of Antimicrobial Chemotherapy, 60, 78-82.
http://dx.doi.org/10.1093/jac/dkm129 [17] Fernando, P., Tania, M., Leonor, G., Melina, R. and Alejandra, C. (2009) Sensitive Screening Tests for Suspected Class A Carbapenemase Production in Species of Enterobacteriaceae. Journal of Clinical Microbiology, 47, 1631-1639.
http://dx.doi.org/10.1128/JCM.00130-09 [18] Poirel, L., Benouda, A., Hays, C. and Nordmann, P. (2011) Emergence of NDM-1 Producing Klebsiella pneumoniae in Morocco. Journal of Antimicrobial Chemotherapy, 66, 2781-2783.
http://dx.doi.org/10.1093/jac/dkr384 [19] Jennifer, C. and Adrian, B. (2011) The Emergence of Carbapenem Resistance in Enterobacteriaceae in South Africa. Southern African Journal of Epidemiology & Infection, 26, 239-240. [20] Rapp, R.P. and Urban, C. (2012) Klebsiella pneumoniae Carbapenemases in Enterobacteriaceae: History, Evolution, and Microbiology Concerns. Pharmacotherapy, 32, 399-407.
http://dx.doi.org/10.1002/j.1875-9114.2012.01035.x [21] Spellberg, B., Blaser, M. and Guidos, R.J. (2011) Combating Antimicrobial Resistance: Policy Recommendations to Save Lives. Clinical Infectious Diseases, 52, 397-405.
http://dx.doi.org/10.1093/cid/cir153
[1] Patrice, N., Laurent, P. and Laurent, D. (2012) Rapid Detection of Carbapenemase Producing Enterobacteriaceae. Emerging Infectious Diseases, 18, 1503-1507.
http://dx.doi.org/10.3201/eid1809.120355 [2] Mammeri, H., Guillon, H.E. and Nordmann, P. (2010) Phenotypic and Biochemical Comparison of the Carbapenems Hydrolyzing Activities of Five Plasmid-Borne AmpC Beta Lactamases. Antimicrobial Agents and Chemotherapy, 54, 4556-4560.
http://dx.doi.org/10.1128/AAC.01762-09 [3] Olaitan, A.O., Berrazeg, M., Fagade, O.E., Adelowo, O.O., Alli, J.A. and Rolain, J.M. (2013) Emergence of Multi Drug Resistant Acinetobacter baumanii Producing OXA-23 Carbapenemase, Nigeria. International Journal of Infectious Diseases, 17, 469-470.
http://dx.doi.org/10.1016/j.ijid.2012.12.008 [4] Centre for Disease Control (2013) Stop Infections from Lethal CRE Germs Now. CDC Vital Signs Report.
http://www.cdc.gov/hai/organisms/cre/TrackingCRE.html [5] Winn, W., Allen, S., Janda, W., Koneman, E., Procop, G., Schreckenberger, P., et al. (2006) Introduction. Koneman’s Color Atlas and Text Book of Diagnostic Microbiology. Lippincott Williams & Wilkins, Baltimore and Philadelphia, 21-25. [6] CLSI (2011) Performance Standards for Antimicrobial Susceptibility Testing: Twenty-First Informational Supplement. CLSI Document M100-S21. Clinical and Laboratory Standard Institute, Wayne. [7] Anderson, K.F., Lonsway, D.R., Rasheed, J.K., Biddle, J., Jensen, B. and McDougal, L.K. (2007) Evaluation of Methods to Identify the Klebsiella pneumoniae Carbapenemase in Enterobacteriaceae. Journal of Clinical Microbiology, 45, 2723-2725.
http://dx.doi.org/10.1128/JCM.00015-07 [8] Van-Tongeren, S.P., Degener, J.E. and Harmsen, H.J.M. (2011) Comparison of Three Rapid and Easy Bacterial DNA Extraction Methods for Use with Quantitative Real Time PCR. European Journal of Clinical Microbiology & Infectious Diseases, 30, 1053-1061.
http://dx.doi.org/10.1007/s10096-011-1191-4 [9] Nordmann, P., Poirel, L., Carrer, A., Toleman, M.A. and Walsh, T.R. (2011) How to Detect NDM-1 Producers. Journal of Clinical Microbiology, 49, 718-721.
http://dx.doi.org/10.1128/JCM.01773-10 [10] Ellington, M.J., Kistler, J., Livermore, D.M. and Woodford, N. (2007) Multiplex PCR for Rapid Detection of Genes Encoding Acquired Metallo-Beta-Lactamases. Journal of Antimicrobial Chemotherapy, 59, 321-322.
http://dx.doi.org/10.1093/jac/dkl481 [11] Pillai, D.R., Melano, R., Rawte, P., Lo, S., Tijet, N., Fuksa, M., et al. (2009) Klebsiella pneumoniae Carbapenemase, Canada. Emerging Infectious Diseases, 15, 827-829.
http://dx.doi.org/10.3201/eid1505.081536 [12] Anna, K.M., Inneke, V.H., Flavia, R., Anna, S.L. and Silvia, F.C. (2009) Multiplex PCR for Rapid Detection of Genes Encoding Oxacillinase and Metallo-Beta-Lactamases in Carbapenem Resistant Acinotobacter Species. JMM Correspondence, 10, 1522-1524.
http://jmm.sgmjournals.org [13] Yusuf, I., Arzai, A.H., Getso, M.I., Sherif, A. and Haruna, M. (2012) Emergence of Carbapenem Resistant Enterobacteriaceae in Surgical and Intensive Care Units of a Hospital with Low Usage of Carbapenem in Kano, North West Nigeria. Antimicrobial Resistance and Infection Control, 2, 75-78.
http://dx.doi.org/10.1186/2047-2994-2-S1-P75 [14] Yusuf, I., Magashi, A.M., Firdausi, F.S., Sharif, A.A., Getso, M.I., Bala, J.A., et al. (2012) Phenotypic Detection of Carbapenemases in Members of Enterobacteriacea in Kano, Nigeria. International Journal of Science and Technology, 2, 802-806. [15] Jones, R.N., Biedenbach, D.J., Sader, H.S., Fritsche, T.R., Toleman, M.A. and Walsh, T.R. (2005) Emerging Epidemic of Metallo-Beta Lactamases Mediated Resistance. Diagnostic Microbiology and Infectious Diseases, 51, 77-84.
http://dx.doi.org/10.1016/j.diagmicrobio.2004.07.003 [16] Landman, D., Bratu, S., Kochar, S., Panwar, M., Trehan, M., Doymaz, M. and Quale, J. (2007) Evolution of Antimicrobial Resistance among Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae in Brooklyn, NY. Journal of Antimicrobial Chemotherapy, 60, 78-82.
http://dx.doi.org/10.1093/jac/dkm129 [17] Fernando, P., Tania, M., Leonor, G., Melina, R. and Alejandra, C. (2009) Sensitive Screening Tests for Suspected Class A Carbapenemase Production in Species of Enterobacteriaceae. Journal of Clinical Microbiology, 47, 1631-1639.
http://dx.doi.org/10.1128/JCM.00130-09 [18] Poirel, L., Benouda, A., Hays, C. and Nordmann, P. (2011) Emergence of NDM-1 Producing Klebsiella pneumoniae in Morocco. Journal of Antimicrobial Chemotherapy, 66, 2781-2783.
http://dx.doi.org/10.1093/jac/dkr384 [19] Jennifer, C. and Adrian, B. (2011) The Emergence of Carbapenem Resistance in Enterobacteriaceae in South Africa. Southern African Journal of Epidemiology & Infection, 26, 239-240. [20] Rapp, R.P. and Urban, C. (2012) Klebsiella pneumoniae Carbapenemases in Enterobacteriaceae: History, Evolution, and Microbiology Concerns. Pharmacotherapy, 32, 399-407.
http://dx.doi.org/10.1002/j.1875-9114.2012.01035.x [21] Spellberg, B., Blaser, M. and Guidos, R.J. (2011) Combating Antimicrobial Resistance: Policy Recommendations to Save Lives. Clinical Infectious Diseases, 52, 397-405.
http://dx.doi.org/10.1093/cid/cir153