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. 2024 Jul 23;16(7):e65185.
doi: 10.7759/cureus.65185. eCollection 2024 Jul.

Phenotypic and Genotypic Characterization of AmpC Beta-Lactamase in Clinical Isolates of Pseudomonas aeruginosa Findings From a Tertiary Care Hospital

Suvarna A Yadav  1 , Satyajeet K Pawar  2 , Kailas D Datkhile  3 , Shivaji T Mohite  4 , Satish R Patil  4 , Ashwini L More  3
Affiliations

Phenotypic and Genotypic Characterization of AmpC Beta-Lactamase in Clinical Isolates of Pseudomonas aeruginosa Findings From a Tertiary Care Hospital

Suvarna A Yadav et al. Cureus. .

Abstract

Background and aim Pseudomonas aeruginosa is an opportunistic pathogen responsible for various healthcare-related infections, which are difficult to treat due to intrinsic and acquired resistance. This study aimed to investigate AmpC β-lactamase production using phenotypic and genotypic methods in Pseudomonas aeruginosa strains isolated from a tertiary care hospital in Karad, Maharashtra, India. Material and methods Over one year, a descriptive cross-sectional study was conducted at the Department of Microbiology, Krishna Institute Medical Sciences, Krishna Vishwa Vidyapeeth, Karad. Phenotypic detection of AmpC beta-lactamase was performed using the Cefoxitin-Cloxacillin Double-Disc Synergy Test method, and genotypic detection was conducted using conventional polymerase chain reaction (PCR) targeting the bla Pseudomonas-derived cephalosporinases (PDC) and bla cephamycinase (CMY) genes. Results Out of 205 clinical isolates of Pseudomonas aeruginosa, 110 (53.66%) showed AmpC production phenotypically, while 86 (41.95%) were positive genotypically. The blaPDC gene was detected in 36.10% of isolates, and the blaCMY gene in 10.73% of isolates. Conclusions The study findings indicate that AmpC-β-lactamase stands out as the primary resistance mechanism in strains of Pseudomonas aeruginosa isolated from the hospital. PCR study concluded that blaPDC (36.10 %) was the leading gene responsible for AmpC synthesis among study isolates. Early detection of AmpC beta-lactamase production by employing phenotypic and genotypic methods is crucial for detecting antibiotic resistance. This dual approach enables healthcare professionals to decide on the most effective antibiotics and mitigate the development of resistance.

Keywords: ampc β-lactamase; antibiotic resistance; blacmy gene; blapdc gene; pseudomonas aeruginosa.

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Conflict of interest statement

Human subjects: Consent was obtained or waived by all participants in this study. INSTITUTIONAL ETHICS COMMITTEE OF KRISHNA INSTITUTE OF MEDICAL SCIENCES KRISHNA INSTITUTE OF MEDICAL SCIENCES "DEEMED TO BE UNIVERSITY", KARAD. issued approval 393/2020-2021. The authors declare that this study was conducted following ethical standards and received approval from the institutional ethics committee of the Krishna Institute of Medical Sciences, Krishna Institute of Medical Sciences “Deemed to be University”, Karad, through protocol number 393/2020-2021. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work.

Figures

Figure 1
Figure 1. Agarose gel image of blaPDC gene having band size 671bp
Lane L was loaded with the 100bp DNA ladder, and Lane 1 with water as a negative control.  Lanes 2,3,4,5 and 7 had positive results, and lane 6 had negative results.
Figure 2
Figure 2. Agarose gel image of blaCMY gene having band size 650bp
Lane L was loaded with the 100bp DNA ladder, lanes 1,3,5 and 7 had positive results, and lanes 2,4, and 6 had negative results.
See this image and copyright information in PMC

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