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Phenotypic and genetic screening of Klebsiella pneumoniae isolates from human UTI patients for beta-lactamases and their genetic diversity analysis by ERIC and REP PCRs

  • Clinical Microbiology - Research Paper
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Abstract

Klebsiella pneumoniae is one of the major nosocomial pathogens responsible for pneumoniae, septicaemia, liver abscesses, and urinary tract infections. Coordinated efforts by antibiotic stewardship and clinicians are underway to curtail the emergence of antibiotic-resistant strains. The objective of the present study is to characterize K. pneumoniae strains through antibiotic resistance screening for production of beta-lactamases (β-lactamases) such as extended spectrum beta lactamases (ESBLs), AmpC β-lactamases, and carbapenemases by phenotypic and genotypic methods and genetic fingerprinting by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and repetitive element palindromic PCR (REP-PCR). A total of 85 K. pneumoniae strains isolated from 504 human urinary tract infections (UTI) were used in this study. Only 76 isolates showed positive in phenotypic screening test (PST), while combination disc method (CDM) as phenotypic confirmatory test (PCT) confirmed 72 isolates as ESBL producers. One or more β-lactamase genes were detected by PCR in 66 isolates (91.66%, 66/72) with blaTEM gene being the most predominant (75.75%, 50/66). AmpC genes could be detected in 21 isolates (31.8%, 21/66) with FOX gene being the predominant (24.24%, 16/66), whereas NDM-I was detected in a single strain (1.51%, 1/66). Genetic fingerprinting using ERIC-PCR and REP-PCR revealed wide heterogeneity among β-lactamase producing isolates with discriminatory power of 0.9995 and 1, respectively.

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Acknowledgements

The authors thank the administration and management of Vignan’s Foundation for Science, Technology and Research (Deemed to be University) as well as NTR College of Veterinary Science, Gannavaram for their support during the study period. PNR thanks Principal, Dr. V.S. Krishna Government Degree College for his encouragement and support during the study period.

Funding

This work was funded by the Department of Science and Technology, Ministry of Science and Technology (grant/award no: SERB/F/4296/2016-17).

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Authors and Affiliations

  1. Department of Biotechnology, Vignan’s Foundation for Science, Technology and Research (Deemed to be University), Vadlamudi, Guntur district, Andhra Pradesh, 522 213, India

    Suresh Bobbadi & Md Nazneen Bobby

  2. Department of Veterinary Public Health and Epidemiology, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, 521101, India

    Bindu Kiranmayi Chinnam

  3. Department of Microbiology, Dr. V.S. Krishna Government Degree and PG College (Autonomous), Maddilapalem, Visakhapatnam, Andhra Pradesh, 530 013, India

    Prakash Narayana Reddy

  4. Division of Veterinary Public Health, ICAR – Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, 243 122, India

    Srinivas Kandhan

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  1. Suresh Bobbadi
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Contributions

SB maintained the isolates and performed the experiments. MNB helped in analysing the data and revising of the manuscript. BKC performed the final revision of the manuscript and approved the final version. PNR is involved in analysing the data, preparation, and final approval of manuscript. SK helped write the initial draft of the manuscript and analyse the data partially.

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Correspondence to Prakash Narayana Reddy.

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Bobbadi, S., Bobby, M.N., Chinnam, B.K. et al. Phenotypic and genetic screening of Klebsiella pneumoniae isolates from human UTI patients for beta-lactamases and their genetic diversity analysis by ERIC and REP PCRs. Braz J Microbiol 54, 1723–1736 (2023). https://doi.org/10.1007/s42770-023-00984-6

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