Extended Spectrum β-Lactamases Producing and Biofilm Forming Multidrug Resistant Klebsiella Pneumoniae from Bacterial Coinfection and Secondary Infection in Patients with Coronavirus Disease At A Tertiary Care Hospital, Kathmandu, Nepal

Authors

  • Supriya Sharma Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal
  • Sushma Regmi Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal
  • Shreedhar Aryal Bhaktapur Hospital, Ministry of Health, Bhaktapur, Nepal
  • Bimal Sharma Chalise Sukraraj Tropical and Infectious Disease Hospital, Teku, Kathmandu, Nepal
  • Krishna Gurung Prithvi Narayan Campus, Tribhuvan University, Pokhara, Nepal
  • Jiwan Thapa Western Regional Hospital, Pokhara, Nepal
  • Sanjib Adhikari Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal
  • Suprina Sharma Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal
  • Komal Raj Rijal Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal
  • Pramod Poudel Central Department of Biotechnology, Tribhuvan University, Kathmandu, Nepal
  • Prakash Ghimire Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal

DOI:

https://doi.org/10.3126/jist.v30i2.82555

Keywords:

Bacterial coinfection, blaCTX-M., ESBL, Klebsiella pneumoniae, Multidrug resistant, Secondary bacterial infection

Abstract

Bacterial co-infections and secondary infections in Coronavirus disease (COVID-19) patients can significantly worsen patient outcomes. Excessive use of antibiotics without prescription during viral infections might result in increase in antimicrobial resistance. This study aimed to determine the prevalence of these bacterial infections and assess multidrug resistance (MDR), Extended Spectrum β -Lactamases (ESBL) production, and biofilm formation among predominant isolates. A prospective study was conducted at Sukraraj Tropical and Infectious Diseases Hospital, Kathmandu, from July 2022 to June 2023. A total of 141 sputum samples from confirmed COVID-19 cases were processed using conventional culture methods. Antimicrobial susceptibility testing was done using the  modified Kirby-Bauer Disc Diffusion method following  Clinical Laboratory Standard Institute guidelines. Biofilm formation was detected by microtitre plate  assay and ESBL production was confirmed phenotypically by the combined disc test. Deoxyribonucleic acid  from ESBL-positive isolates was tested for the blaCTX-M gene using conventional polymerase chain reaction.  Out of 141 patients,  9 (6.38%) showed bacterial co-infections and 11 (7.80%) showed secondary bacterial infections. Gram-negative bacteria were predominant, with Klebsiella pneumoniae 12 (60.00%) being the most common isolate. High resistance rates were observed in K. pneumoniae for Ceftriaxone 8 (66.70%) and Amikacin 7 (58.30%), while Imipenem remained effective for 11 (91.7%) isolates. Among the bacterial isolates  14/20 (70.00%) were MDR, 8/20 (40.00%) were biofilm producers and 8/19 (42.10%) were ESBL producers, with 6/8 (75.00%) harboring the blaCTX-M gene. To the best of our knowledge, this is one of the few studies to report ESBL producing MDR bacteria isolated from co-infection and secondary bacterial infection among COVID positive cases in Nepal. Continuous monitoring of bacterial infections and resistance patterns is crucial for improving infection control and managing the pandemic.

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References

Abrar, S., Ain, N. U., Liaqat, H., Hussain, S., Rasheed, F., & Riaz, S. (2019). Distribution of blaCTX − M, blaTEM, blaSHV and blaOXA genes in Extended-spectrum-β-lactamase-producing Clinical isolates: A three-year multi-center study from Lahore, Pakistan. Antimicrobial Resistance & Infection Control, 8(1), 80. https://doi.org/10.1186/s13756-019-0536-0.

Acharya, M., Joshi, P. R., Thapa, K., Aryal, R., Kakshapati, T., & Sharma, S. (2017). Detection of metallo-β-lactamases-encoding genes among clinical isolates of Pseudomonas aeruginosa in a tertiary care hospital, Kathmandu, Nepal. BMC Research Notes, 10(1), 718. https://doi.org/10.1186/s13104-017-3068-9.

Alkhouli, M., Nanjundappa, A., Annie, F., Bates, M. C., & Bhatt, D. L. (2020). Sex Differences in Case Fatality Rate of COVID-19: Insights From a Multinational Registry. Mayo Clinic Proceedings, 95(8), 1613–1620. https://doi.org/10.1016/j.mayocp.2020.05.014.

Clancy, C. J., & Nguyen, M. H. (2020). Coronavirus Disease 2019, Superinfections, and Antimicrobial Development: What Can We Expect? Clinical Infectious Diseases, 71(10), 2736–2743. https://doi.org/10.1093/cid/ciaa524.

CLSI. (2020). Performance Standards for Antimicrobial Susceptibility Testing. Clinical and Laboratory Standards Institute. https://clsi.org.

Devkota, S. P., Sharma, S., Bhatta, D. R., Paudel, A., Sah, A. K., & Kandel, B. P. (2018). Prevalence of the blaNDM gene among metallo-β-lactamase-producing Gram-negative isolates from western Nepal. Journal of Global Antimicrobial Resistance, 12, 3–4. https://doi.org/10.1016/j.jgar.2017.11.003.

Dhungana, K., Krishna Awal, B., Dhungel, B., Sharma, S., Raj Banjara, M., & Raj Rijal, K. (2019). Detection of Klebsiella pneumoniae Carbapenemase (KPC) and Metallo-Beta Lactamase (MBL) Producing Gram Negative Bacteria Isolated from Different Clinical Samples in A Transplant Center, Kathmandu, Nepal. Acta Scientific Microbiology, 2(12), 60–69. https://doi.org/10.31080/ASMI.2019.02.0432.

Duan, Y., Wang, J., Wang, S., Zhang, R., Hu, J., Li, W., & Chen, B. (2024). Risk factors, outcomes, and epidemiological and etiological study of hospitalized COVID-19 patients with bacterial co-infection and secondary infections. European Journal of Clinical Microbiology & Infectious Diseases, 43(3), 577–586. https://doi.org/10.1007/s10096-024-04755-5.

Karataş, M., Yaşar-Duman, M., Tünger, A., Çilli, F., Aydemir, Ş., & Özenci, V. (2021). Secondary bacterial infections and antimicrobial resistance in COVID-19: Comparative evaluation of pre-pandemic and pandemic-era, a retrospective single center study. Annals of Clinical Microbiology and Antimicrobials, 20(1), 51. https://doi.org/10.1186/s12941-021-00454-7.

Khurana, S., Singh, P., Sharad, N., Kiro, V. V., Rastogi, N., Lathwal, A., Malhotra, R., Trikha, A., & Mathur, P. (2021). Profile of co-infections & secondary infections in COVID-19 patients at a dedicated COVID-19 facility of a tertiary care Indian hospital: Implication on antimicrobial resistance. Indian Journal of Medical Microbiology, 39(2), 147–153. https://doi.org/10.1016/j.ijmmb.2020.10.014.

Kot, B. (2019). Antibiotic Resistance Among Uropathogenic Escherichia coli. Polish Journal of Microbiology, 68(4), 403–415. https://doi.org/10.33073/pjm-2019-048.

Lansbury, L., Lim, B., Baskaran, V., & Lim, W. S. (2020). Co-infections in people with COVID-19: A systematic review and meta-analysis. Journal of Infection, 81(2), 266–275. https://doi.org/10.1016/j.jinf.2020.05.046.

Liu, Y., Ling, L., Wong, S. H., Wang, M. H., Fitzgerald, J. R., Zou, X., Fang, S., Liu, X., Wang, X., Hu, W., Chan, H., Wang, Y., Huang, D., Li, Q., Wong, W. T., Choi, G., Zou, H., Hui, D. S., Yu, J., … Zhang, L. (2021). Outcomes of respiratory viral-bacterial co-infection in adult hospitalized patients. eClinicalMedicine, 37, 100955. https://doi.org/10.1016/j.eclinm.2021.100955.

Maharjan, R., Bastola, A., Adhikari, N., Rijal, K. R., Banjara, M. R., Ghimire, P., & Shrestha, U. T. (2022). Multidrug-resistant bacteria with ESBL genes: A growing threat among people living with HIV/AIDS in Nepal. BMC Infectious Diseases, 22(1), 526. https://doi.org/10.1186/s12879-022-07503-2.

Mutua, J. M., Njeru, J. M., & Musyoki, A. M. (2022). Multidrug resistant bacterial infections in etseverely ill COVID-19 patients admitted in a national referral and teaching hospital, Kenya. BMC Infectious Diseases, 22(1), 877. https://doi.org/10.1186/s12879-022-07885-3.

Nepal, R., Shrestha, B., Joshi, D. M., Joshi, R. D., Shrestha, S., & Singh, A. (2018). Antibiotic Susceptibility Pattern of Gram-negative Isolates of Lower Respiratory Tract Infection. Journal of Nepal Health Research Council, 16(1), 22–26. https://doi.org/10.3126/jnhrc.v16i1.19358.

Nirwati, H., Sinanjung, K., Fahrunissa, F., Wijaya, F., Napitupulu, S., Hati, V. P., Hakim, M. S., Meliala, A., Aman, A. T., & Nuryastuti, T. (2019). Biofilm formation and antibiotic resistance of Klebsiella pneumoniae isolated from clinical samples in a tertiary care hospital, Klaten, Indonesia. BMC Proceedings, 13(S11), 20. https://doi.org/10.1186/s12919-019-0176-7.

Parajuli, N. P., Maharjan, P., Joshi, G., & Khanal, P. R. (2016). Emerging Perils of Extended Spectrum β -Lactamase Producing Enterobacteriaceae Clinical Isolates in a Teaching Hospital of Nepal. BioMed Research International, 2016, 1–7. https://doi.org/10.1155/2016/1782835.

Saeed, N. K., Al-Khawaja, S., Alsalman, J., Almusawi, S., Albalooshi, N. A., & Al-Biltagi, M. (2021). Bacterial co-infection in patients with SARS-CoV-2 in the Kingdom of Bahrain. World Journal of Virology, 10(4), 168–181. https://doi.org/10.5501/wjv.v10.i4.168.

Shakya, G., Shrestha, S., Adhikari, S., Rijal, N., & Sharma, S. (n.d.). Changing trend of antimicrobial susceptibility pattern of Salmonella spp. In different regions of Nepal: A six years surveillance study.

Sharifipour, E., Shams, S., Esmkhani, M., Khodadadi, J., Fotouhi-Ardakani, R., Koohpaei, A., Doosti, Z., & Ej Golzari, S. (2020). Evaluation of bacterial co-infections of the respiratory tract in COVID-19 patients admitted to ICU. BMC Infectious Diseases, 20(1), 646. https://doi.org/10.1186/s12879-020-05374-z.

Shrestha, S. D., Malla, S., Adhikari, B., Shakya, G., Basnyat, S., & Sharma, S. (2010). Antibiotic Susceptibility Patterns of Vibrio cholerae isolates. Journal of Nepal Medical Association, 49(179). https://doi.org/10.31729/jnma.95.

Tille, P. M. (2020). BAILEY & SCOTT’S DIAGNOSTIC MICROBIOLOGY. CHURCHILL LIVINGSTONE.

Yang, X., Li, X., Qiu, S., Liu, C., Chen, S., Xia, H., Zeng, Y., Shi, L., Chen, J., Zheng, J., Yang, S., Tian, G., Liu, G., & Yang, L. (2024). Global antimicrobial resistance and antibiotic use in COVID-19 patients within health facilities: A systematic review and meta-analysis of aggregated participant data. Journal of Infection, 106183. https://doi.org/10.1016/j.jinf.2024.106183.

Zhou, F., Yu, T., Du, R., Fan, G., Liu, Y., Liu, Z., Xiang, J., Wang, Y., Song, B., Gu, X., Guan, L., Wei, Y., Li, H., Wu, X., Xu, J., Tu, S., Zhang, Y., Chen, H., & Cao, B. (2020). Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: A retrospective cohort study. The Lancet, 395(10229), 1054–1062. https://doi.org/10.1016/S0140-6736(20)30566-3.

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Published

2025-12-23

How to Cite

Sharma, S., Regmi, S., Aryal, S., Sharma Chalise, B., Gurung, K., Thapa, J., … Ghimire, P. (2025). Extended Spectrum β-Lactamases Producing and Biofilm Forming Multidrug Resistant Klebsiella Pneumoniae from Bacterial Coinfection and Secondary Infection in Patients with Coronavirus Disease At A Tertiary Care Hospital, Kathmandu, Nepal. Journal of Institute of Science and Technology, 30(2), 105–114. https://doi.org/10.3126/jist.v30i2.82555

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Research Articles