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Background <p>Hospital-acquired infections (HAIs) associated with increasing antimicrobial resistance (AMR) are becoming a global health challenge, causing AMR spread and a serious health threat. This study was designed to evaluate the phenotypic and genotypic status of AMR in Gram-negative HAI-associated bacteria, to gain a current insight into the local settings.</p> Methods and Results <p>Kirby-Bauer method and Double Disc Synergy Test were used for phenotypic assessment of AMR and extended-spectrum β-lactamase (ESBL) producers, respectively. Genes encoding antimicrobial resistance, ESBLs, and carbapenemases were identified by PCR followed by Sanger sequencing. Among 102 HAI samples, 62 were Gram-negative, identified as <i>Escherichia coli</i> (<i>E. coli</i>) (42.2%), <i>Klebsiella pneumoniae</i> (<i>K. pneumoniae</i>) (21.1%), and <i>Pseudomonas aeruginosa</i> (<i>P. aeruginosa</i>) (6.6%) by PCR. All <i>K. pneumoniae</i> and (95%) <i>E. coli</i> isolates exhibited multidrug resistance. <i>K. pneumoniae</i> isolates (95%) showed resistance against ceftazidime, cefotaxime, ceftriaxone, and aztreonam, and (89%) isolates were resistant to amoxicillin-clavulanic acid, ofloxacin, and cefepime (<i>E. coli</i> isolates (84%), (84%), (78%), (76%), (76%), and (73%) were resistant against cefotaxime, ceftriaxone, aztreonam, ceftazidime, doripenem, and colistin, respectively. Genotypically, the <i>mcr-2</i> gene was detected in 5% <i>K. pneumoniae</i> isolates. Phenotypically, 63% isolates were identified as ESBL producers with high genetic prevalence of SHV-12 (78.9%) and SHV-1 (57.8%) in <i>K. pneumoniae</i> and CTX-M1 (60.5%) in <i>E. coli.</i> Notably, one <i>K. pneumoniae</i> isolate harbored all class-A ESBL genes, which is a novel finding. Co-existence of CTX-M1 + SHV-1 + TEM-1, SHV-1 + SHV-12 + TEM-1, and CTX-M1 + CTX-M15 + TEM-1 genes was dominant.</p> Conclusion <p>The HAIs caused by extensively drug-resistant and ESBL-producing <i>K. pneumoniae</i> and <i>E. coli</i> were dominant, with less prevalence of moderately drug-resistant <i>Pseudomonas</i> isolates.</p>

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High prevalence and co-existence of SHV and CTX-M genes in extensively drug-resistant (XDR) Gram-negative bacteria isolated from hospital settings in Faisalabad District, Pakistan

  • 期刊论文

  • 发表日期：

    2026年3月13日

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  • Umair Abbas,
  • Huma Sajjad,
  • Amir Shaukat,
  • Zaheer Ahmad,
  • Azfar Sattar,
  • Khurram Shehzad,
  • Mark A. T. Blaskovich,
  • Tania Da Silva Duarte,
  • Johannes Zuegg,
  • Aamir Ali,
  • Yasra Sarwar

摘要

Background

Hospital-acquired infections (HAIs) associated with increasing antimicrobial resistance (AMR) are becoming a global health challenge, causing AMR spread and a serious health threat. This study was designed to evaluate the phenotypic and genotypic status of AMR in Gram-negative HAI-associated bacteria, to gain a current insight into the local settings.

Methods and Results

Kirby-Bauer method and Double Disc Synergy Test were used for phenotypic assessment of AMR and extended-spectrum β-lactamase (ESBL) producers, respectively. Genes encoding antimicrobial resistance, ESBLs, and carbapenemases were identified by PCR followed by Sanger sequencing. Among 102 HAI samples, 62 were Gram-negative, identified as Escherichia coli (E. coli) (42.2%), Klebsiella pneumoniae (K. pneumoniae) (21.1%), and Pseudomonas aeruginosa (P. aeruginosa) (6.6%) by PCR. All K. pneumoniae and (95%) E. coli isolates exhibited multidrug resistance. K. pneumoniae isolates (95%) showed resistance against ceftazidime, cefotaxime, ceftriaxone, and aztreonam, and (89%) isolates were resistant to amoxicillin-clavulanic acid, ofloxacin, and cefepime (E. coli isolates (84%), (84%), (78%), (76%), (76%), and (73%) were resistant against cefotaxime, ceftriaxone, aztreonam, ceftazidime, doripenem, and colistin, respectively. Genotypically, the mcr-2 gene was detected in 5% K. pneumoniae isolates. Phenotypically, 63% isolates were identified as ESBL producers with high genetic prevalence of SHV-12 (78.9%) and SHV-1 (57.8%) in K. pneumoniae and CTX-M1 (60.5%) in E. coli. Notably, one K. pneumoniae isolate harbored all class-A ESBL genes, which is a novel finding. Co-existence of CTX-M1 + SHV-1 + TEM-1, SHV-1 + SHV-12 + TEM-1, and CTX-M1 + CTX-M15 + TEM-1 genes was dominant.

Conclusion

The HAIs caused by extensively drug-resistant and ESBL-producing K. pneumoniae and E. coli were dominant, with less prevalence of moderately drug-resistant Pseudomonas isolates.