Molecular Characterization of Extended Spectrum Beta-lactamase from Enterobacter cloacae, E. coli and Klebsiella pneumoniae from Pregnant Women in South-south Nigeria
International Journal of Microbiology and Biotechnology
Volume 5, Issue 2, June 2020, Pages: 48-54
Received: Feb. 7, 2020; Accepted: Feb. 21, 2020; Published: Apr. 21, 2020
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Florence Zion Uyanga, Department of Microbiology, Akwa Ibom State University, Mkpat Enin, Nigeria
Emmanuel Olufemi Ekundayo, Departments of Microbiology, Michael Okpara University of Agriculture, Umudike, Nigeria
Emmanuel Onwubiko Nwankwo, Departments of Microbiology, Michael Okpara University of Agriculture, Umudike, Nigeria
Inimfon Akaninyene Ibanga, Department of Microbiology, Akwa Ibom State University, Mkpat Enin, Nigeria
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Extended spectrum beta-lactamase prevalence is still on the increase across the world and has been implicated in urinary tract infections in Nigeria. TEM, SHV and CTX-M are becoming more common with CTX-M-15 becoming more significant as it is associated with complicated urinary tract infections. This study was conducted to evaluate distribution of blaTEM, blaCTX-M-15 and blaSHV genes among Enterobacter cloacae, E. coli and Klebsiella pneumoniae. A total of 660 urine samples were collected from pregnant women in 3 general hospitals from Akwa Ibom state. Enterobacter cloacae, E. coli and Klebsiella pneumoniae were identified using Microbact 24E. The disc diffusion and combined discs methods were used for testing antimicrobial susceptibility. The presence of ESBL was detected using Double Disk Synergy Test (DDST) and CHROMagar ESBL, respectively. Plasmid extraction was carried out following the protocol of ZR Plasmid Miniprep-Classic extraction kit. The blaCTX-M-15, blaTEM and blaSHV was identified by PCR with specific primers in selected 50 ESBL producing isolates. A total of 252 clinical isolates were collected from three General Hospitals in South-South, Nigeria. ESBLs were found in 231 (92%) isolates. blaCTX-M-15 was the commonest genotype (84%), followed by blaSHV (60%) and blaTEM (60%). ESBL positive strains of E. cloacae, E. coli and K pneumoniae are increasingly found in isolates from pregnant women. The current study demonstrated the predominance of gene encoding blaCTX-M-15 with a percentage frequency of (84%) commonly with plasmid in the setting.
ESBL blagene, PCR, UTI
To cite this article
Florence Zion Uyanga, Emmanuel Olufemi Ekundayo, Emmanuel Onwubiko Nwankwo, Inimfon Akaninyene Ibanga, Molecular Characterization of Extended Spectrum Beta-lactamase from Enterobacter cloacae, E. coli and Klebsiella pneumoniae from Pregnant Women in South-south Nigeria, International Journal of Microbiology and Biotechnology. Vol. 5, No. 2, 2020, pp. 48-54. doi: 10.11648/j.ijmb.20200502.12
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Paterson, D. L. and Bonomo, R. A. (2005) Extended-spectrum β-lactamases: A Clinical update. Clin Microbiol Rev. 18: 657-686.
Shashwati, N., Kiran, T. and Dhanvijay, A. (2014) Study of Extended Spectrum β-lactamase Producing Enterobacteriaceae and Antibiotic co-resistance in a Tertiary Care Teaching Hospital. J Nat Sci Biol Med. 5: 30-35.
Ma, L., Lin, C., Chen, J., Fung, C., Chang, F. et al.(2009) and the Taiwan Surveillance of Antimicrobial Resistance Project (2009) Widespread dissemination of aminoglycoside resistance genes armA and rmtB in Klebsiella pneumoniae isolates in Taiwan producing CTX-M-type extended-spectrum β-lactamases. Antimicrob Agents Chemoth. 53: 104-111.
Sharma, M., Pathak, S. and Srivastava, P. (2013) Prevalence and Antibiogram of Extended Spectrum β-lactamase (ESBL) producing Gram negative bacilli and further molecular Characterization of ESBL producing Escherichia coli and Klebsiella spp. J Clin Diagn Res. 7: 2173-2177.
Bradford, P. A. (2001) Extended-spectrum β-lactamases in the 21st century: Characterization, Epidemiology, and Detection of this important Resistance Threat. Clin Microbiol Rev. 14: 933-951.
Kalp, M., Bethel, C. R., Bonomo, R. A. and Carey, P. R. (2009) Why the Extended-spectrum β-lactamases SHV-2 and SHV-5 are “Hyper susceptible” to Mechanism-based inhibitors. Biochemistry. 48: 9912-9920.
Sullivan, R., Schaus., D, John, M. and Delport, J. A. (2015). Extended spectrum beta-lactamases: A minireview of clinical relevant groups. J Med Microb Diagn.; 4 (203): 2161-703.1000203.
Bonnedahl, J., Hernandez, J., Stedt, J., Waldenström, J., Olsen, B. et al. (2014) Extended-spectrum β-lactamases in Escherichia coli and Klebsiella pneumoniae in gulls, alaska, USA. Emerg Infect Dis. 20: 897-899.
Bauernfeind, A., Casellas, J. M., Goldberg, M., Holley, M., Jungwirth, R., Mangold, P., Röhnisch, T., Schweighart, S. and Wilhelm, R. (1992). A new plasmidic cefotaximase from patients infected with Salmonella typhimurium. 20 (3): 158-63.
Bauernfeind, A., Grimm, H., Schweighart, S. (1990). A new Plasmidic Cefotaximase in a Clinical Isolate of Escherichia coli Infection 1990; 18; 294-8.
Bernard, H., Tancrede. C., Livrelli, V. et al. (1992). A Novel plasmid-mediated Extended-spectrum β-lactamase not derived from TEM-or SHV-type enzymes. J Antimicrob Chemother 1992; 29: 590–2.
Bevan, E. R., Jones, A. M. and Hawkey, P. M. (2017). Global Epidemiology of CTX-M β-lactamases: Temporal and geographical shifts in genotype. J. Antimicrob. Chemother. 72, 2145–2155.
Woerther, P. L., Burdet, C., Chachaty, E. et al. Trends in Human Fecal Carriage of Extended-spectrum β-lactamases in the Community: Goward the globalization of CTX-M. Clin Microbiol Rev, 2013; 26: 744—58.
CDC/HICPAC, (2009). Urinary Tract Infection (UTI) Event for Long-Term Care Facilities, CDC, Atlanta, GA, USA.
Roopa, T. J. and Sudha, S. S. (2010). Antimicrobial susceptibility of extended spectrum beta –lactamase (ESBL) producing uropathogens isolated from ICU patients. Int J Biological Technology. 1: 23–31.
Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing: Twenty-first informational supplement. CLSI 2011 Document M100-S21.
Jarlier, V., Nicolas, M. H., Fournier, G. and Philippon A. (1988). ESBLs conferring transferable resistance to newer-lactam agents in Enterobacteriaceae: Hospital prevalence and susceptibility patterns. Rev Infect Dis. 10 (4): 867–878.
Chaudhary, U. and Aggarwal, R. (2004). Extended spectrum beta lactamase: An emerging threat to clinical therapeutics. Ind J Med Microbiol 22 (2): 75-80.
Ho, P. L., Tsang, D. N., Que T. L., Ho, M., and Yuenk Y.(2000). Comparison of screening Methods for Detection of Extended Spectrum Beta lactamase and their Prevalence among E. coli and Klebsiella spp in Hong Kong. APMIS. 108 (3): 237-240.
Samyyia, Abrar., Shahida, Hussain., Rehan, A. K., Noor, U. l., Ain, H, H. and Saba, R.(2018). Prevalence of Extended-spectrum-β-lactamase-producing Enterobacteriaceae: First systematic meta-analysis report from Pakistan. Antimicrobial Resistance and Infection Control, 7; 26.
Baguma, A, Atek, K. and Joel, B. (2017). Prevalence of Extended-Spectrum Beta-Lactamases-Producing Microorganisms in Patients Admitted at KRRH, Southwestern Uganda, International Journal of Microbiology, ID 3183076, 5 pages.
Mutasim, E. I, Mohammed, A., Abdullah, M., A. and Bahaeldin K. E. (2019). Phenotypic Characterization and Antibiotic Resistance Patterns of Extended-Spectrum β-Lactamase-and AmpC β-Lactamase-Producing Gram-Negative Bacteria in a Referral Hospital, Saudi Arabia, Hindawi, Canadian Journal of Infectious Diseases and Medical Microbiology, Article ID 6054694, 9 pages,
Marie, M. A., John, J. Krishnappa, L. G. and Gopalkrishnan, S. (2013) “Molecular Characterization of the β-lactamases in Escherichia coli and Klebsiella pneumoniae from a Tertiary care Hospital in Riyadh, Saudi Arabia,” Microbiology and Immunology, vol. 57, no. 12, pp. 805–810.
Olowo-Okere, A. Y., Ibrahim, K. E. and Olayinka, B. O. (2018) “Molecular Characterisation of ESBL producing Gram-negative Bacteria isolates from Surgical Wounds of Patients at a Hospital in North Central Nigeria,” Journal of Global Antimicrobial Resistance, vol. 14, pp. 85–89.
Habeeb, M. A. Sarwar, Y. Ali, A. Salman, M. and Haque, A. (2013) “Rapid emergence of ESBL producers in E. coli causing Urinary and Wound Infections in Pakistan,” Pakistan Journal of Medical Sciences, vol. 29, no. 2, pp. 540–544.
Dalela, G. (2012). Prevalence of extended spectrum beta-lactamase (ESBL) producers among gram-negative bacilli from various Clinical Isolates in a Tertiary care Hospital at Jhalawar, Rajasthan, India. J Clin Diagn Res. 6 (2): 182–7.
Östholm-Balkhed, Å., Tärnberg, M., Nilsson, M. et al., (2010). “Prevalence of Extended-spectrum beta-lactamase-producing Enterobacteriaceae and Trends in Antibiotic Consumption in a County of Sweden,” Scandinavian Journal of Infectious Diseases, vol. 42, no. 11-12, pp. 831–838, 2010.
Willemsen, I. Oome, S. Verhulst, C. Pettersson, A. Verduin, V and Kluytmans, J. (2015).“Trends in extended spectrum beta-lactamase (ESBL) producing Enterobacteriaceae and ESBL genes in a Dutch teaching Hospital, measured in 5 yearly point Prevalence Surveys (2010–2014),” PLoS One, vol. 10, no. 11, Article ID e0141765, 10 pages.
Somily, A. M., Habib, H. A., Absar, M. M., et al.,(2014) “ESBL-producing Escherichia coli and Klebsiella pneumoniae at a Tertiary Care Hospital in Saudi Arabia,” Journal of Infection in Developing Countries, vol. 8, no. 9, pp. 1129–1136, 2014.
Kobra, S. Rizi, 1., Shahin, N. Peerayeh, 1., Bita, B. and Mohammad, R. (2015). Emergence of blaCTX-M-15 Gene and Its Transferability in Enterobacter spp. Isolated From the Hospitals of Tehran, Iran Avicenna Clin Microb Infect. 2 (3) e29691.
Mirsalehian, A., Akbari, N. F., Peymani, A., Kazemi, B., Jabal, A., Mirafshar, S. M.(2008) Prevalence of Extended Spectrum Beta-lactamase-producing Enterobacteriaceae by Phenotypic and Genotypic methods in Intensive care Units in Tehran, Iran. Daru. 16 (3): 169–73.
Kasap, M., Fashae, K., Torol, S., Kolayli, F., Budak, F, Vahaboglu, H. (2010). Characterization of ESBL (SHV-12) producing Clinical Isolate of Enterobacter aerogenes from a Tertiary care Hospital in Nigeria. Ann Clin Microbiol Antimicrob. 2010; 9: 1.
Morand, P. C., Billoet, A., Rottman, M,., Sivadon-Tardy, V., Eyrolle, L., Jeanne, L., et al. (2009) Specific distribution within the Enterobacter cloacae complex of strains isolated from infected orthopedic Implants. J Clin Microbiol. 2009; 47 (8): 2489–95.
Huang, Z. M., Shan, H., Mi, Z. H., Yang, H. Y., Wu, L., Chu, Q. J., et al. (2008). [Analysis on 168 rRNA methylase Genes and Aminoglycoside modifying Enzymes Genes in Enterobacter cloacae in China]. Zhonghua Liu Xing Bing Xue Za Zhi. 29 (4): 369–73.
Iabadene, H., Messai, Y., Ammari, H., Ramdani-Bouguessa, N., Lounes, S. Bakour, R. et al. (2008). Dissemination of ESBL and Qnr determinants, in Enterobacter cloacae in Algeria. J Antimicrob Chemother. 62 (1): 133–6.
Ifeanyi, A. O. and Florence E. Orok (2015). The Bacterial Isolates and Plasmid Profile of Extended Spectrum Beta-Lactamases Producers Causing Urinary Tract Infection among Pregnant Women in Uyo, Nigeria Journal of Biosciences and Medicines, 2015, 3, 25-30.
Abrar, S., Vajeeha, A., Ul-Ain, N., Riaz, S. (2017). Distribution of CTX-M group I and group III β-lactamases produced by Escherichia coli and Klebsiella pneumoniae in Lahore, Pakistan. Microb Pathog. 103: 8–12.
Sonda, T., Kumburu, H., Van Zwetselaar, M., Alifrangis, M., Mmbaga, B. T., Lund, O. et al. (2018). Prevalence and risk factors for CTX-M Gram-negative bacteria in Hospitalized patients at a Tertiary care Hospital in Kilimanjaro, Tanzania. Eur J Clin Microbiol Infect Dis. 37: 1–10.
Zhao, D., Quan, J., Liu, L., Du, X., Chen, Y., Jiang, Y., et al. (2016). High prevalence of ESBL-producing Escherichia coli and Klebsiella pneumoniae in community-onset bloodstream infections in China. J Antimicrob Chemother. 72 (1): 273–80.
Pavez, M., Troncoso, C., Osses, I., Salazar, R., Illesca, V., Reydet, P, et al.(2019) High prevalence of CTX-M-1 group in ESBL-producing Enterobacteriaceae Infection in Intensive care units in Southern Chile. Braz J Infect Dis. 53: 25-30.
Doi, Y., Park, Y. S., Rivera, J. I., Adams-Haduch, J. M., Hingwe, A., Sordillo, E. M., Lewis, J. S., 2nd, Howard, W. J., Johnson, L. E., Polsky, B., Jorgensen, J. H., Richter, S. S., Shutt, K. A. and Paterson, D. L. (2013) Community-associated extended-spectrum beta-lactamase-producing Escherichia coli infection in the United States. Clin Infect Dis. 56 (5): 641–648.
Livermore, D. M., Canton, R., Gniadkowski, M., Nordnman, P., Rossolin, G. M. Arlet, G. (2007). et al. CTX-M: Changing the face of ESBLs in Europe. J Antimicrob Chemother. 59: 165-74.
Sid Ahmed, M. A., Bansal, D., Acharya, A., Elmi, A. A., Hamid, J. M., Sid Ahmed, A. M. et al.(2016) Antimicrobial susceptibility and molecular epidemiology of extended-spectrum beta-lactamase-producing Enterobacteriaceae from intensive care units at Hamad Medical Corporation, Qatar. Antimicrob Resist Infect Control. 5 (1): 4–9.
Vaida, S., Mafius, L., Aurelija, B., Justas, P., Rita, P., Agnes, G. et al., (2010). Molecular Characteristics of extended-spectrum beta-lactamase producing E. coli and K. pneumoniae Isolates from Hospitals in Lithuania, Journal of Medical Microbiology 59 (10): 1263-65.
Kotekani, L., Kotigadde, S., (2018). Virulence determinant and extended spectrum beta-lactamase production in Klebsiella pneumoniae isolated from a Tertiary care Hospital, South India. J Lab Physicians 10: 155-61.
Wang, G., Huang, T., Surendraiah, P. K., Wang, K., Komal, R., Zhuge J. et al (2013). CTX-M β-lactamase-producing Klebsiella pneumoniae in suburban New York city, New York, USA. Emerg Infect Dis19: 1803-1808.
Mshana, S. E., Hain, T., Domann, E., Lyamuya, E. F., Chakraborty, T., Imirzalioglu, C, (2013). Predominance of Klebsiella pneumoniae ST14 carrying CTX-M-15 causing neonatal sepsis in Tanzania. BMC Infect Dis; 13: 466.
Hashemi A. (2014) Detection of blactx-m, blatem, blashv genes in Klebsiella pneumoniae strains isolated from two hospitals of Tehran, Iran. Iran, J Public Health. 43 (2): 98.
Ahmad, H. P. and Khalil, M. K. (2019). Prevalence of blaTEM, blaSHV, and blaCTX-M Genes among ESBL-Producing Klebsiella pneumoniae and Escherichia coli Isolated from Thalassemia Patients in Erbil, Iraq, Mediterr J Hematol Infect Dis 11 (1): e2019041.
Xia, S., Fan, X., Huang, Z., Xia, L., Xiao M, Chen R, et al. (2014) Dominance of CTX-M-type extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolated from patients with community-onset and hospital-onset infection in China. PLoS One.; 9 (7): e100707.
Shivakumara, S., Karthik, P., Bhagya, T., Ranjan, D., Rani, B., Bhavani, M, Niranjana, M., Bulagonda, E. P. (2018). Characterization of blaCTX-M Sequences of Indian origin and thirteen uropathogenic Escherichia coli isolates Resistant to multiple antibiotics BMC Res Notes (2018) 11: 630.
Mazzariol, A., Bazaj, A. and Cornaglia G. (2017) Multi-drug-resistant Gram-negative Bacteria causing Urinary Tract Infections: A Review. J Chemother; 29 (sup1): 2-9.
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