Molecular characterization and transmission pattern of tetracycline resistance gene in tigecycline and carbapenem resistance Klebsiella pneumoniae isolates at a tertiary care hospital India

2026 · doi:10.1099/acmi.0.001017.v3
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This study examined carbapenem- and tigecycline-resistant Klebsiella pneumoniae from pus and respiratory clinical specimens at a tertiary care hospital in India, using identification by biochemical tests and MALDI-TOF MS, AST by disc diffusion with MICs by broth microdilution, and PCR/Sanger sequencing to detect carbapenemase genes (blaNDM, blaOXA-48, blaKPC) and tetracycline resistance genes (tet(A), tet(B), tet(K), tet(M), tet(S)). Among 152 carbapenem-resistant isolates, 20.4% (31) were resistant to tigecycline and showed high multidrug resistance, and blaOXA-48 (51.6%) and blaNDM (29.0%) were present with blaKPC not detected; tet(A) (12.9%) and tet(B) (3.2%) were found, while tet(K), tet(M), and tet(S) were not. Conjugation experiments showed transferable plasmids carrying blaNDM and blaOXA-48, whereas plasmids with tet(A) and tet(B) were not transferable under the experimental conditions. The paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.

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Abstract

Abstract: Background: The increasing prevalence of tigecycline and carbapenem-resistant K. pneumoniae (CRKP) poses a serious challenge, especially in resource-limited settings. Its ability to exchange resistance genes with other bacteria accelerates the spread of multidrug resistance. While carbapenems and tetracyclines have been used effectively against K. pneumoniae, resistance to these agents is now rising globally, narrowing available treatment options. Objective: The study aimed to determine the phenotypic and genotypic prevalence of carbapenem and tetracycline resistance K. pneumoniae isolates along with transferability pattern of carbapenem and tetracycline resistance gene in these isolates. Methodology Clinical isolates from pus and respiratory samples were identified using biochemical tests and MALDI-TOF MS. AST was performed by the Kirby-Bauer disc diffusion method, and MICs were determined by BMD method. PCR was performed to detect carbapenemase (blaNDM, blaOXA-48, blaKPC) and tetracycline resistance genes [tet(A), tet(B), tet(K), tet(M), tet(S)], followed by Sanger sequencing for validation. Conjugation assays assessed gene transferability. Results Out of 152 carbapenem resistant K. pneumoniae isolates, 31 (20.4%) were found to be resistant to tigecycline. All tigecycline-resistant isolates (31/31; 100%) exhibited complete resistance to ceftazidime, ciprofloxacin, and omadacycline. Additionally, resistance to amikacin and cefoperazone-sulbactam (SCF) was observed in 87.1% (27/31) and 77.4% (24/31) of the isolates. Resistance to minocycline and colistin was detected in 51.6% (16/31) and 29.0% (9/31) of the isolates, respectively. PCR analysis revealed that 51.6% (16/31) of the isolates carried the blaOXA-48 gene, and 29.0% (9/31) carried the blaNDM gene. None of the isolates harbored the blaKPC gene. With respect to tetracycline resistance determinants, the tet(A) gene was detected in 12.9% (4/31) of the isolates, and the tet(B) gene in 3.2% (1/31), while tet(K), tet(M), tet(S) and blaKPC were not detected in any isolate. Conjugation assays demonstrated that plasmids carrying blaNDM and blaOXA-48 were transferable to a recipient strain, indicating their potential for horizontal gene transfer. In contrast, plasmids harboring tet(A) and tet(B) genes were not transferable under the experimental conditions. Conclusion Tigecycline-resistant K. pneumoniae isolates showed high multidrug resistance, with transferable blaNDM and blaOXA-48 genes. In contrast, Chromosome and plasmid borne tetracycline resistance genes tet(A) and tet(B) were non-transferable, indicating limited horizontal spread.
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Abstract

Background: The increasing prevalence of tigecycline and carbapenem-resistant K. pneumoniae (CRKP) poses a serious challenge, especially in resource-limited settings. Its ability to exchange resistance genes with other bacteria accelerates the spread of multidrug resistance. While carbapenems and tetracyclines have been used effectively against K. pneumoniae, resistance to these agents is now rising globally, narrowing available treatment options.

Objective

The study aimed to determine the phenotypic and genotypic prevalence of carbapenem and tetracycline resistance K. pneumoniae isolates along with transferability pattern of carbapenem and tetracycline resistance gene in these isolates. Methodology: Clinical isolates from pus and respiratory samples were identified using biochemical tests and MALDI-TOF MS. AST was performed by the Kirby-Bauer disc diffusion method, and MICs were determined by BMD method. PCR was performed to detect carbapenemase (blaNDM, blaOXA-48, blaKPC) and tetracycline resistance genes [tet(A), tet(B), tet(K), tet(M), tet(S)], followed by Sanger sequencing for validation. Conjugation assays assessed gene transferability.

Results

Out of 152 carbapenem resistant K. pneumoniae isolates, 31 (20.4%) were found to be resistant to tigecycline. All tigecycline-resistant isolates (31/31; 100%) exhibited complete resistance to ceftazidime, ciprofloxacin, and omadacycline. Additionally, resistance to amikacin and cefoperazone-sulbactam (SCF) was observed in 87.1% (27/31) and 77.4% (24/31) of the isolates. Resistance to minocycline and colistin was detected in 51.6% (16/31) and 29.0% (9/31) of the isolates, respectively. PCR analysis revealed that 51.6% (16/31) of the isolates carried the blaOXA-48 gene, and 29.0% (9/31) carried the blaNDM gene. None of the isolates harbored the blaKPC gene. With respect to tetracycline resistance determinants, the tet(A) gene was detected in 12.9% (4/31) of the isolates, and the tet(B) gene in 3.2% (1/31), while tet(K), tet(M), tet(S) and blaKPC were not detected in any isolate. Conjugation assays demonstrated that plasmids carrying blaNDM and blaOXA-48 were transferable to a recipient strain, indicating their potential for horizontal gene transfer. In contrast, plasmids harboring tet(A) and tet(B) genes were not transferable under the experimental conditions.

Conclusion

Tigecycline-resistant K. pneumoniae isolates showed high multidrug resistance, with transferable blaNDM and blaOXA-48 genes. In contrast, Chromosome and plasmid borne tetracycline resistance genes tet(A) and tet(B) were non-transferable, indicating limited horizontal spread. - Received: - Version Posted:

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