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CC BY-NC-ND 4.0 · J Lab Physicians 2019; 11(03): 253-258
DOI: 10.4103/JLP.JLP_31_19
Original Article

Detection and genetic characterization of extended-spectrum beta-lactamases producers in a tertiary care hospital

Suryarashmi Sahoo
Department of Microbiology, Institute of Medical Sciences and SUM Hospital, S ‘O’ A University, Bhubaneswar, Odisha, India
,
Sarita Otta
Department of Microbiology, Institute of Medical Sciences and SUM Hospital, S ‘O’ A University, Bhubaneswar, Odisha, India
,
Bichitrananda Swain
Department of Microbiology, Institute of Medical Sciences and SUM Hospital, S ‘O’ A University, Bhubaneswar, Odisha, India
,
Subrat Kumar Kar
Regional Plant Resource Centre, Bhubaneswar, Odisha, India
› Author Affiliations Financial support and sponsorship The study was financially supported by S’O’A University, Kalinga Nagar, Bhubaneswar, Odisha.
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Abstract

BACKGROUND: Extended-spectrum beta-lactamase (ESBL)-producing organisms inactivate extended beta-lactam antibiotics and monobactams and also exhibit coresistance to many other classes of antibiotics. The present study was carried out to assess the prevalence of the ESBLs and to determine the most prevalent genotype in our hospital.

MATERIALS AND METHODS: All clinically significant Gram-negative isolates were identified, and their antimicrobial susceptibility testing was done by Kirby–Bauers' disc diffusion method. ESBL detection was confirmed by minimal inhibitory concentration method using agar dilution technique for those who screened positive by ceftazidime (30 μg) disc. Further, the established ESBL-positive isolates were subjected to genotyping for bla TEM, bla CTX-M, and bla SHV genes by using conventional polymerase chain reaction.

RESULTS: Escherichia coli was the most common (28.84%) Gram-negative bacillus followed by Klebsiella pneumoniae (18.07%), while Pseudomonas spp. (9.61%) was the most commonly identified nonfermenter. ESBL production was detected in 160 (30.8%) isolates. Klebsiella oxytoca (46.7%) followed by E. coli (44%) were the common ESBL producers. Most predominant ESBL gene was bla TEM, found in 122 (76.25%) isolates. Combinations of two genes were seen in 109 (68.1%) isolates, the most common (43.12%) combination being blaTEM and blaCTX-M. In this study, 16 (10%) strains had all the three types of genes. Most of the isolated Gram-negative bacilli (GNB) were sensitive to amikacin, imipenem, and colistin.

CONCLUSION: In our study, the 30.8% of GNB were ESBL producers. This is the only study that shows that TEM is the most prevalent ESBL genotypes in our area. Of concern is a good number of isolates showing all three patterns of genes (TEM, SHV, and CTX-M). Amikacin, imipenem, and colistin were the most useful antibiotics in our setup.

Key words

Bla TEM - extended-spectrum beta-lactamase - genotyping


Publication History

Received: 20 February 2019

Accepted: 16 August 2019

Article published online:
07 April 2020

© 2019.

Thieme Medical and Scientific Publishers Private Ltd.
A-12, Second Floor, Sector -2, NOIDA -201301, India

 

  • References

  • 1 Wiegand I, Hilpert K, Hancock RE. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc 2008;3:163-75.
  • 2 Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 1989.
  • 3 Yano H, Kuga A, Irinoda K, Okamoto R, Kobayashi T, Inoue M. Presence of genes for beta-lactamases of two different classes on a single plasmid from a clinical isolate of Serratia marcescens. J Antibiot (Tokyo) 1999;52:1135-9.
  • 4 Batchelor M, Hopkins K, Threlfall EJ, Clifton-Hadley FA, Stallwood AD, Davies RH, et al. Bla (CTX-M) genes in clinical Salmonella isolates recovered from humans in England and wales from 1992 to 2003. Antimicrob Agents Chemother 2005;49:1319-22.
  • 5 Vandana KE, Honnavar P. Amp C beta-lactamase among ESBL-producing E. coli and If you don't look, you won't find. J Clin Diagn Res 2009;3:1635-56.
  • 6 Winokur PL, Canton R, Casellas JM, Legakis N. Variations in the prevalence of strains expressing an extended-spectrum beta-lactamase phenotype and characterization of isolates from Europe, the Americas, and the Western Pacific region. Clin Infect Dis 2001;32 Suppl 2:S94-103.
  • 7 Bouchillon SK, Johnson BM, Hoban DJ, Johnson JL, Dowzicky MJ, Wu DH, et al. Determining incidence of extended spectrum beta-lactamase producing Enterobacteriaceae, vancomycin-resistant Enterococcus faecium and methicillin-resistant Staphylococcus aureus in 38 centres from 17 countries: The PEARLS study 2001-2002. Int J Antimicrob Agents 2004;24:119-24.
  • 8 Mathai D, Manoharan A, Vasanthan G. Epidemiology and Implications of ESBL. Crit Care Update 2009;14:152-62.
  • 9 Fouzia B, Damle AS. Genotypic detection of extended-spectrum β-lactamase-producing Klebsiella pneumoniae in a Tertiary care hospital. Int J Biomed Adv Res 2015;6:91-7.
  • 10 Jena J, Debata NK, Subudhi E. Prevalence of extended-spectrum-beta-lactamase and metallo-beta-lactamase producing multi drug resistance gram – Negative bacteria from urinary isolates. Indian J Med Microbiol 2013;31:420-1.
  • 11 Livermore DM, Hawkey PM. CTX-M: Changing the face of ESBLs in the UK. J Antimicrob Chemother 2005;56:451-4.
  • 12 Babypadmini S, Appalaraju B. Extended spectrum -lactamases in urinary isolates of Escherichia coli and Klebsiella pneumoniae – Prevalence and susceptibility pattern in a tertiary care hospital. Indian J Med Microbiol 2004;22:172-4.
  • 13 Laudy AE, Róg P, Smolińska-Król K, Ćmiel M, Słoczyńska A, Patzer J, et al. Prevalence of ESBL-producing Pseudomonas aeruginosa isolates in Warsaw, Poland, detected by various phenotypic and genotypic methods. PLoS One 2017;12:e0180121.
  • 14 Sinha M, Srinivasa H, Macaden R. Antibiotic resistance profile and extended spectrum beta-lactamase (ESBL) production in Acinetobacter species. Indian J Med Res 2007;126:63-7.
  • 15 Joshi SG, Litake GM, Ghole VS, Niphadkar KB. Plasmid-borne extended-spectrum beta-lactamase in a clinical isolate of Acinetobacter baumannii. J Med Microbiol 2003;52:1125-7.
  • 16 Yong D, Shin JH, Kim S, Lim Y, Yum JH, Lee K, et al. High prevalence of PER-1 extended-spectrum beta-lactamase-producing Acinetobacter spp. In Korea. Antimicrob Agents Chemother 2003;47:1749-51.
  • 17 Ansari S, Nepal HP, Goutam R, Shrestha J, Neopane P, Gurung G, et al., Community acquired multi-drug resistant clinical isolates of Escherichia coli in a tertiary care centre of Nepal. Antimicrob Resist Infect Control 2015;4:15.
  • 18 Sasirekha B, Manasa R, Ramya P, Sneha R. Frequency and antimicrobial sensitivity pattern of extended spectrum β-lactamases producing Escherichia coli and Klebsiella pneumoniae isolated in a tertiary care hospital. Al Ameen J Med Sci 2010;3:265-71.
  • 19 Manoharan A, Premalatha K, Chatterjee S, Mathai D; SARI Study Group. Correlation of TEM, SHV and CTX-M extended-spectrum beta lactamases among Enterobacteriaceae with their in vitro antimicrobial susceptibility. Indian J Med Microbiol 2011;29:161-4.
  • 20 Rahman MM, Haq JA, Hossain MA, Sultana R, Islam F, Islam AH. Prevalence of extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in an urban hospital in Dhaka, Bangladesh. Int J Antimicrob Agents 2004;24:508-10.
  • 21 Kaur M, Aggarwal A. Occurrence of the CTX-M, SHV and the TEM genes among the extended spectrum β-lactamase producing isolates of Enterobacteriaceae in a tertiary care hospital of North India. J Clin Diagn Res 2013;7:642-5.
  • 22 Rezai MS, Salehifar E, Rafiei A, Langaee T, Rafati M, Shafahi K, et al. Characterization of multidrug resistant extended-spectrum beta-lactamase-producing Escherichia coli among uropathogens of pediatrics in North of Iran. Biomed Res Int 2015;2015:309478.
  • 23 Bajpai T, Pandey M, Varma M, Bhatambare GS. Prevalence of TEM, SHV, and CTX-M beta-lactamase genes in the urinary isolates of a tertiary care hospital. Avicenna J Med 2017;7:12-6.
  • 24 Shahid M, Singh A, Sobia F, Rashid M, Malik A, Shukla I, et al. Bla (CTX-M), Bla (TEM), and Bla (SHV) in Enterobacteriaceae from North-Indian tertiary hospital: High occurrence of combination genes. Asian Pac J Trop Med 2011;4:101-5.
  • 25 Livermore DM, Sefton AM, Scott GM. Properties and potential of ertapenem. J Antimicrob Chemother 2003;52:331-44.