Efficacy of colistin-based combinations against pandrug-resistant whole-genome-sequenced Klebsiella pneumoniae isolated from hospitalized patients in Egypt: an in vitro/vivo comparative study DOI Creative Commons
Eriny T. Attalla, Amal M Khalil, Azza S. Zakaria

et al.

Gut Pathogens, Journal Year: 2024, Volume and Issue: 16(1)

Published: Dec. 3, 2024

Abstract Background Colistin resistance significantly constrains available treatment options and results in the emergence of pandrug-resistant (PDR) strains. Treating PDR infections is a major public health issue. A promising solution lies using colistin-based combinations. Despite availability vitro data evaluating these combinations, vivo studies remain limited. Results Thirty colistin-resistant Klebsiella pneumoniae (ColRKp) isolates were collected from hospitalized patients. was detected broth microdilution, antimicrobial susceptibility tested Kirby-Bauer method against 18 antibiotics. Extremely high levels detected, with 17% being PDR. Virulence profiling, assessed Anthony capsule staining, string test, crystal violet assay, indicated predominance non-biofilm formers non-hypermucoid The screened for mcr genes polymerase chain reaction. Whole-genome sequencing (WGS) bioinformatics analysis performed to characterize genomes isolates. No plasmid-borne WGS revealed that belonged high-risk clones: ST14 (n = 1), ST147 2), ST383 2). They carried encoding extended-spectrum β-lactamases carbapenemases, bla CTX-M-15 NDM-5 , on conjugative IncHI1B/IncFIB plasmids, illustrating convergence virulence genes. most common mechanism colistin involved alterations mgrB . Furthermore, deleterious amino acid substitutions also within PhoQ, PmrC, CrrB, ArnB, ArnT. Seven colistin-containing combinations compared checkerboard experiment. Synergy observed when combining tigecycline, doxycycline, levofloxacin, ciprofloxacin, sulfamethoxazole/trimethoprim, imipenem, or meropenem. efficacy combined either doxycycline levofloxacin modulation vivo, murine infection model. In vitro, reversed 80% 73.3% population, respectively. + combination demonstrated superiority over rescuing infected animals, reducing bacterial bioburden liver kidneys while preserving nearly intact lung histology. Conclusions This study represents first comparative investigation clinical ColRKp characterized at genomic level.

Language: Английский

Exploring AMR and virulence in Klebsiella Pneumoniae isolated from humans and pet animals: A complement of phenotype by WGS-derived profiles in a one-health study in Egypt DOI Creative Commons

Enas Soliman,

Alaa Saad,

Ashraf Abd El-Tawab

et al.

One Health, Journal Year: 2024, Volume and Issue: 19, P. 100904 - 100904

Published: Sept. 27, 2024

Language: Английский

Citations

6
Assessment of Phenotypic Tools for Detection of OXA-48, KPC, and NDM in Klebsiella pneumoniae in Oman DOI Creative Commons

Arwa AL Rujaibi,

Zaaima AL-Jabri,

Amina Al Jardani

et al.

Diagnostics, Journal Year: 2025, Volume and Issue: 15(8), P. 949 - 949

Published: April 8, 2025

Background: The alarming increase in carbapenemase-producing Enterobacterales is a matter of grave public health concern. most ubiquitous carbapenemases, Klebsiella pneumoniae carbapenemase (KPC)-, New Delhi metallo-β-lactamase (NDM)-, and oxacillinase (OXA-48)-like enzymes, belong to the Ambler molecular classes A, B, D, respectively. KPC- OXA-48-like enzymes have serine-based hydrolytic mechanism, while NDMs are metallo-β-lactamases that contain zinc active site. For judicious use reserve drugs promoting antimicrobial stewardship, timely detection carbapenemases essential. While tools gold standard for these many laboratories limited access them. This study focused on evaluating in-house commercial phenotypic tests OXA-48-, KPC-, NDM-like K. pneumoniae, predominant extremely drug-resistant pathogen Oman. Methods: In total, 80 GeneXpert/PCR-confirmed (40 OXA-48 20 KPC NDM each) 37 whole-genome-sequenced (25 OXA-232 6 KPC-2, plus NDM-1 NDM-5) were subjected screening by temocillin (30 μg disk) (MAST Diagnostica, Germany) D71C (MASTDISCS®). Isolates resistant (<11 mm) four tests: an tool (OXA-48 disk test) three (i) MASTDISCS® Combi (D72C) Group Ltd., Bootle, UK); (ii) (D73C) (iii) immunochromatographic assay (ICT), which KPC/IMP/NDM/VIM/OXA-48 Combo test kit (Medomics, China), carbapenemases. Results: Temocillin exhibited good sensitivity specificity (100% 97.50%) compared (70% 100%). Among confirmatory tests, had 92.50% 100% specificity, MAST DISC D72C, D73C, ICT 97.50%, 95.00%, 100%, 91.67%, 95% Conclusions: tool. With high ease performance, short turnaround time, low cost, we recommend format routine diagnostic use. resource-constrained centers, excellent alternative with specificity.

Language: Английский

Citations

0
Genome Characterization of Carbapenem-Resistant Hypervirulent Klebsiella pneumoniae Strains, Carrying Hybrid Resistance-Virulence IncHI1B/FIB Plasmids, Isolated from an Egyptian Pediatric ICU DOI Creative Commons

Heba Ali Hammad,

Radwa Abdelwahab,

Douglas F. Browning

et al.

Microorganisms, Journal Year: 2025, Volume and Issue: 13(5), P. 1058 - 1058

Published: May 1, 2025

Despite the increased reporting of Carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKp) in Egypt, there is a paucity information regarding molecular characteristics such strains. Herein, we present genome sequence two CR-hvKp strains, K22 and K45, which were isolated from VAP (ventilator-associated-pneumonia) patients admitted to pediatric ICU at Assiut University Children’s Hospital, Egypt. K45 isolates subjected antimicrobial susceptibility testing whole-genome sequencing. Genomic analysis was performed characterize each strain, determining their plasmids, resistance (AMR) genes, virulence determinants. possessed an extensive drug phenotype (XDR), whilst exhibited multidrug (MDR), with sequencing revealing presence diverse array AMR genes. Both strains resistant carbapenem antibiotic imipenem, carrying OXA-48 carbapenemase, additionally possessing NDM-1 carbapenemase. Each strain considered high-risk, respectively belonging types ST383 ST14 genes implicated hypervirulence (e.g., iucABCD-iutA rmpA). Importantly, both carried multiple plasmid replicons, including AMR/virulence IncHI1B/FIB hybrid MDR IncL/M plasmids. This report highlights critical role plasmids evolution virulent K. suggests circulation plasmid, simultaneously disseminating hypervirulence, amongst within Hospital.

Language: Английский

Citations

0
A novel XbaI multiplex PCR method for rapid typing of Klebsiella pneumoniae strains DOI Creative Commons
Ibrahim Alzahrani

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: April 26, 2025

This study focused on exploiting single-nucleotide polymorphism (SNP) variations in and around XbaI-restriction sites (5'….T↓CTAGA….3') within the genomes of Klebsiella pneumoniae to develop a multiplex PCR genotyping technique that integrates high discrimination pulsed-field gel electrophoresis (PFGE) with straightforwardness PCR-based procedure, for routine use clinical laboratories. The XbaI-multiplex method was evaluated silico using 10 reference strains subsequently compared PFGE MLST, revealing similar clustering patterns test most cases. Furthermore, 29 isolates known sequence types (STs) were analysed ed PCR, which demonstrated acceptable discriminative ability relative concordance MLST results. Like other typing methods, is not only cost-effective but also user-friendly as it does necessitate complex instruments or specialized skills, results are available 4-6 h. Its implementation relies standard laboratory instruments, such machine agarose electrophoresis. However, still requires further evaluations larger number isolates, including K. obtained from hospital outbreaks.

Language: Английский

Citations

0
Molecular detection of OXA-48 and NDM-1 carbapenemase genes among clinical isolates of Klebsiella pneumoniae recovered from patients attending a private tertiary hospital in Southwestern Nigeria DOI Creative Commons
Chisom Blossom Onyeji, Seyi Samson Enitan, Olalekan Ademola Kemiki

et al.

BMC Infectious Diseases, Journal Year: 2024, Volume and Issue: 24(1)

Published: Sept. 13, 2024

Language: Английский

Citations

0
Efficacy of colistin-based combinations against pandrug-resistant whole-genome-sequenced Klebsiella pneumoniae isolated from hospitalized patients in Egypt: an in vitro/vivo comparative study DOI Creative Commons
Eriny T. Attalla, Amal M Khalil, Azza S. Zakaria

et al.

Gut Pathogens, Journal Year: 2024, Volume and Issue: 16(1)

Published: Dec. 3, 2024

Abstract Background Colistin resistance significantly constrains available treatment options and results in the emergence of pandrug-resistant (PDR) strains. Treating PDR infections is a major public health issue. A promising solution lies using colistin-based combinations. Despite availability vitro data evaluating these combinations, vivo studies remain limited. Results Thirty colistin-resistant Klebsiella pneumoniae (ColRKp) isolates were collected from hospitalized patients. was detected broth microdilution, antimicrobial susceptibility tested Kirby-Bauer method against 18 antibiotics. Extremely high levels detected, with 17% being PDR. Virulence profiling, assessed Anthony capsule staining, string test, crystal violet assay, indicated predominance non-biofilm formers non-hypermucoid The screened for mcr genes polymerase chain reaction. Whole-genome sequencing (WGS) bioinformatics analysis performed to characterize genomes isolates. No plasmid-borne WGS revealed that belonged high-risk clones: ST14 (n = 1), ST147 2), ST383 2). They carried encoding extended-spectrum β-lactamases carbapenemases, bla CTX-M-15 NDM-5 , on conjugative IncHI1B/IncFIB plasmids, illustrating convergence virulence genes. most common mechanism colistin involved alterations mgrB . Furthermore, deleterious amino acid substitutions also within PhoQ, PmrC, CrrB, ArnB, ArnT. Seven colistin-containing combinations compared checkerboard experiment. Synergy observed when combining tigecycline, doxycycline, levofloxacin, ciprofloxacin, sulfamethoxazole/trimethoprim, imipenem, or meropenem. efficacy combined either doxycycline levofloxacin modulation vivo, murine infection model. In vitro, reversed 80% 73.3% population, respectively. + combination demonstrated superiority over rescuing infected animals, reducing bacterial bioburden liver kidneys while preserving nearly intact lung histology. Conclusions This study represents first comparative investigation clinical ColRKp characterized at genomic level.

Language: Английский

Citations

0