Search Results (355)

Emerging antimicrobial resistance (AMR) in companion animals represents a global health concern as they serve as potential reservoirs for multidrug-resistant (MDR) bacteria, which can be transmitted to humans. Herein, we provide comprehensive surveillance data on resistance patterns in veterinary hospital settings, focusing on urinary tract infection. A total of 23 bacterial strains were isolated from urine specimens of hospitalized companion animals suspected of urinary tract infections (UTIs) between 2022 and 2024. 16S rRNA sequencing analysis revealed that Escherichia coli (47.8%), Klebsiella pneumoniae (21.7%), and Pseudomonas aeruginosa (8.7%) were predominant uropathogens. Minimum inhibitory concentration and minimum bactericidal concentration tests were employed to analyze AMR patterns across different classes of antibiotics. Moreover, antimicrobial susceptibility test exhibited 73.91% MDR according to the standard definition given by the Clinical and Laboratory Standards Institute (CLSI) M100 guidelines. Most Gram-negative bacteria have been shown to be resistant to beta-lactam antibiotics, especially carbapenems. Notably, an E. coli strain was confirmed to possess the bla_NDM-1 gene encoding the carbapenemase New Delhi metallo-β-lactamase. These findings support the implementation of targeted infection control measures and evidence-based treatment protocols to preserve antimicrobial efficacy in companion animal medicine to minimize potential public health risks through the One Health approach. Full article

Background/Objectives: Antimicrobial resistance (AMR) among Enterobacterales poses a major threat to public health in Southern Africa and has led to limited treatment options and increased mortality. Despite Africa bearing the brunt, there is limited data on the epidemiology and molecular epidemiology of the genetic determinants of β-lactam and/or carbapenem resistance. This narrative literature review summarizes the epidemiology and molecular characteristics of extended-spectrum β-lactamase-producing Enterobacterales (ESBL-PE), carbapenem-resistant Enterobacterales (CRE), and carbapenemase-producing Enterobacterales (CPE) in Southern Africa, while identifying data gaps and surveillance challenges. Methods: A comprehensive literature review was conducted using peer-reviewed articles from ten Southern African countries, including South Africa, Lesotho, Eswatini, Botswana, Namibia, Angola, Zambia, Zimbabwe, Mozambique, and Malawi, reporting the epidemiology and/or molecular characterization of ESBL-PE, CRE, and CPE. Results: ESBL-PE, CRE, and CPE pose an increasing healthcare threat in Southern Africa, with prevalence varying widely by source. Klebsiella pneumoniae and E. coli are the predominant ESBL-PE, CRE, and CPE species. The most frequent resistance genes are bla_CTX-M among ESBLs and bla_NDM and bla_OXA among carbapenemases, reflecting global patterns. However, molecular characterization across the region remains limited, with countries such as Botswana, Lesotho, Eswatini, Zambia, and Zimbabwe lacking sufficient data on the prevalence and diversity of these resistance determinants. Conclusions: Despite the paucity of genomic and epidemiological data, Southern Africa faces an urgent AMR challenge. Strengthening laboratory infrastructure, genomic surveillance, and regional coordination is crucial to mitigate AMR and guide antibiotic stewardship policies. Full article

The skin of turtles, particularly aquatic species, can harbor a diverse range of bacteria, including Citrobacter species, which are recognized as causative agents of Septicemic Cutaneous Ulcerative Disease. Consequently, turtles may act as reservoirs of pathogenic and multidrug-resistant bacteria, posing a potential public health concern. This case-based study investigated the presence of Citrobacter spp. in a loggerhead sea turtle (Caretta caretta) housed at the Livorno Aquarium, Italy. Nine swabs were collected from skin lesions (plastron, carapace, nuchal mass), the oral cavity, and the cloaca. The isolated strains were identified by MALDI-TOF MS and tested for their susceptibility to 12 antimicrobials, belonging to eight antimicrobial classes, by the disc diffusion method. Isolates were investigated genotypically for extended-spectrum-β-lactamase (ESBL) bla_CTX−M, bla_TEM, bla_SHV, bla_PER, and metallo-β-lactamase (MBL) bla_IMP, bla_OXA−48, bla_VIM, bla_NDM, bla_GES genes. Biofilm production ability was also evaluated. Fifteen Citrobacter spp. strains were recovered from the analyzed samples. Complete resistance was recorded for ampicillin, followed by high levels of resistance to imipenem, tetracycline and piperacillin-tazobactam. Worryingly, 86.7% were classified as multidrug-resistant. The most common ESBL-genotype combination was bla_SHV and bla_PER genes (60%), while the most frequently detected MBL gene was bla_NDM (46.7%), followed by bla_GES (40%). Most isolates were classified as weak biofilm producers (80%). The findings of this study demonstrate the presence of Citrobacter spp., an opportunistic pathogen, with a notable prevalence of multidrug-resistant strains carrying beta-lactamase-encoding genes, in a loggerhead sea turtle in Italy, across both lesioned and healthy anatomical sites. Full article

It is crucial to identify Enterobacterales that produce carbapenemase to treat and manage hospital infections. The suggested techniques for their identification need a lengthy wait, technical knowledge, and training. Lateral flow immunoassays (LFIAs) provide a solution to these requirements. Thus, this study compared LFIA with phenotypic and genotypic tests for carbapenemase-producing bacteria. Fifty clinical isolates of carbapenem-resistant superbugs were examined. KPC, VIM, NDM, IMP, and OXA-48-like enzymes were evaluated and compared with phenotypic tests and LFIA. Regarding the phenotypic characteristics, the mCIM was positive in 37/50 (74%), and the eCIM was positive in 21/50 (42%). Regarding using LFIA, 41 out of the total isolates (82%) gave a positive red line with one or more of the tested genes. The most frequently detected gene was bla_NDM (27/50 (54%)), and the least detected one was bla_IMP (14/50 (28%)), which was in accordance with the PCR results. While investigating the accuracy of LFIA vs. PCR, it was found that LFIA had 100% sensitivity in the detection of the bla_NDM and bla_OXA genes, with 85.2% and 91.4% specificity, respectively, while for the bla_IMP, bla_KPC, and bla_VIM genes, the values were 91.7% and 92.1%, 94.1% and 90.9%, and 95.5% and 89.3%, respectively. The overall accuracy of LFIA ranged from 92 to 94%. Our comparison with molecular assays revealed remarkable agreement, so we propose that this test might be utilized as a supplementary tool. Full article

Background/Objectives: The environmental dissemination of antimicrobial-resistant Enterobacteriaceae poses a remarkable threat to public health. This study investigates the environmental presence and dissemination of carbapenemase-producing Enterobacteriaceae (CPE) in 30 important water bodies selected according to their interconnection with and utilization by livestock and community people in central Thailand. Methods: Water samples were collected from 30 selected water bodies. Enterobacteriaceae were isolated and screened for CPE and multidrug resistance. Carbapenemase genes (bla_NDM-5, bla_NDM-1 and bla_IMI-1) were detected and their locations (plasmid and chromosome) determined. Plasmid types were further characterized, and conjugation experiments were performed to assess transferability among bacterial species. Results: From all selected samples, six isolates (20%) were identified as multidrug-resistant CPE including one Escherichia coli, one Klebsiella pneumoniae and four Enterobacter roggenkampii carrying bla_NDM-5, bla_NDM-1 and bla_IMI-1 genes, respectively. The bla_NDM-5 and bla_NDM-1 genes were located on phage-like pO111 type plasmid and IncC plasmid, while bla_IMI-1 was located on chromosomes. The plasmids also consisted of components that closely resembled those found in resistance plasmids obtained from clinical and environmental isolates worldwide. Additionally, through plasmid conjugation experiment, carbapenemase genes were transferable with a high rate among bacterial species. Conclusions: These findings indicated that water bodies are polluted and there is an urgent need for integrated strategies to monitor and mitigate the spread of antibiotic resistance across human, animal and environmental health domains in aquatic environments. Full article

Escherichia coli sequence type 648 (ST648), a lineage within the clinically important phylogroup F, has disseminated worldwide in humans and animals. In this study, we performed whole-genome sequencing and comparative genomic analysis for two New Delhi metallo-beta-lactamase (bla_NDM-5) carrying E. coli strains: ECsOL198, recovered from a wild Eurasian otter in Northern Lebanon, and ECOL247, isolated from a hospitalized leukemia patient. Both isolates belonged to phylogroup F and serotype O9:H4, and exhibited IncFIA, IncFIB, and IncFII plasmids. They shared a similar antimicrobial resistance profile, including a carbapenemase gene (bla_NDM-5), β-lactamase genes (bla_TEM-1, bla_CTX-M-15, and bla_OXA-1), and other genes that confer resistance to aminoglycosides (acc(3)-Ile, aadA2), sulfonamides (sul1), tetracyclines (tet(A)), and fluoroquinolones (mutations in gyrA and parC). Both isolates also carried common virulence-associated genes related to adhesion, iron acquisition, environmental persistence, and immune evasion. Whole-genome multilocus sequence typing (wgMLST) revealed that both isolates formed a distinct subclade closely related to a bloodstream-derived ST648 isolate from India, indicating limited relatedness to global clones. These findings highlight the transmission of nearly clonal multidrug-resistant E. coli ST648 in both clinical and non-clinical settings, raising concerns about the threat to public health. Full article

Background: Rapid antimicrobial susceptibility testing (RAST) allows early detection of resistance directly from positive blood cultures, potentially improving outcomes in bloodstream infections (BSIs). We evaluated the performance of EUCAST RAST for Gram-negative ESKAPEEc pathogens and characterized carbapenemase genes in carbapenem-resistant Klebsiella pneumoniae (CRKP). Methods: A total of 354 positive blood cultures were screened, including 51 monomicrobial Gram-negative ESKAPEEc isolates. RAST results at 4, 6, 8, and 16–20 h were compared with standard antimicrobial susceptibility testing (AST) obtained using the BD Phoenix™ system. Categorical agreement (CA) and error frequency were calculated. Multiplex PCR and Sanger sequencing were performed on 15 CRKP isolates to identify carbapenemase genes and allelic variants. Results: 51 Gram-negative ESKAPEEc isolates met the inclusion criteria for RAST (15 E. coli, 19 K. pneumoniae, 11 A. baumannii, and 6 P. aeruginosa). Overall performance varied markedly by species and antibiotic. E. coli showed frequent unreadable or ATU zones at early timepoints and wide CA variability (50–100%), with high very major error (VME) rates for AMP, TZP, and CAZ, particularly at 6–8 h. K. pneumoniae displayed consistently high CA (mostly 100%) for carbapenems, CAZ, and TZP. A. baumannii demonstrated excellent agreement (100% for most agents), except for GEN at 6–8 h. P. aeruginosa could be evaluated only at 16–20 h, showing high CA for AMK, CAZ, and CIP; lower CA for MEM (83%); non-calculable CA for IMI due to universal ATU readings; and a CA value of 0% for TZP due to the predominance of the ATU results. VMEs ranged from 0% to 26.1% across species and reading times, but carbapenems did not generate VMEs. Molecular analysis revealed bla_KPC in 66.7%, bla_NDM in 46.7%, and bla_OXA-48 in 33.3% of isolates, with co-occurrence in several strains. Sequencing identified bla_KPC-2 and bla_NDM-1 as the predominant variants, with one isolate harboring bla_NDM-5. Conclusions: EUCAST RAST markedly accelerates susceptibility reporting from positive blood cultures, but its accuracy is species- and time-dependent. Performance was excellent for K. pneumoniae (including CRKP) and A. baumannii and acceptable for P. aeruginosa at 16–20 h. In contrast, E. coli showed frequent ATU results at early timepoints and high ME/VME rates, making readings before 8 h unreliable for clinical decisions. Overall, RAST can effectively support rapid antimicrobial stewardship when species-specific limitations are recognized, and early-timepoint results are interpreted with caution. Full article

Background/Objectives: Providencia stuartii is intrinsically resistant to several antibiotic classes, and acquisition of bla_NDM further restricts treatment options. This study aimed to characterize NDM-producing P. stuartii isolates from a small hospital cluster in Russia and to place them within the global genomic context. Methods: Four isolates recovered between June and July 2023 from a single hospital were analyzed using Illumina and Oxford Nanopore sequencing to assess genetic relatedness and plasmid content. Results: The isolates showed identical extensively drug-resistant profiles and were closely related genomically. All carried nearly identical IncC plasmids harboring multiple antimicrobial resistance genes, including bla_NDM. Comparative analysis indicated that these genomes clustered with recent European isolates but differed in the bla_NDM allele and its genomic location. Highly similar IncC plasmids were also found in our previous Klebsiella pneumoniae dataset, demonstrating that this plasmid backbone occurs in multiple bacterial species in the region. Conclusions: The study highlights the role of IncC plasmids in carbapenemase dissemination and underscores the value of genomic surveillance integrating chromosomal and plasmid analyses to track extensively drug-resistant pathogens. Full article

Background/Objectives: The global emergence of carbapenem resistance is a major public health concern. Campylobacter jejuni and Campylobacter coli, key zoonotic agents causing human campylobacteriosis, are mainly isolated from poultry, their primary host. Their increasing resistance in animals and humans highlights the risk of gene transfer. This study investigates the molecular mechanisms of carbapenem resistance in 287 avian Campylobacter spp. isolates from Tunisia within a One Health approach. Methods: Antibiotic susceptibility of 287 carbapenem-resistant isolates, including 147 C. jejuni and 140 C. coli, was determined according to CLSI. All isolates were screened by PCR for genes encoding the most reported carbapenemases, including VIM, IMP, NDM and OXA-48. Eleven multidrug-resistant (MDR)/carbapenem-resistant C. coli isolates were selected to determine their clonal lineage by Multilocus sequence typing (MLST). Results: All isolates were susceptible to imipenem, but resistance to meropenem and ertapenem were observed in 60.71% and 35.71% of C. coli isolates, respectively, versus 13.6% in C. jejuni for each antibiotic. The bla_VIM, bla_NDM and bla_OXA-48 genes were detected in 15, 8, and 19 of the 20 C. jejuni isolates, respectively. However, for C. coli, 53, 12, and 15 isolates harbored bla_VIM, bla_NDM and bla_OXA-48 genes, respectively. The eleven (MDR)/carbapenem-resistant C. coli isolates belonged to a unique ST sequence type ST13450. Conclusions: We report for the first time the emergence of bla_VIM, bla_NDM, and bla_OXA-48 genes in Campylobacter spp. isolates of poultry origin highlighting possible horizontal transfer of these genes to pathogenic Gram-negative bacteria of the poultry’s microbiota. Full article

Carbapenem-resistant Klebsiella pneumoniae isolates harbouring double carbapenemases, from patients in a surgical and transplantation ICU, were investigated to better understand the dispersion of the pathogen. Twenty-three carbapenem-resistant K. pneumoniae isolates harbouring at least two different carbapenemases (by immunochromatography screening), were consecutively collected during a seven-month period from patients in a surgical and transplantation ICU. Identification and susceptibility testing were performed using the MALDI-TOF Vitek MS and the Vitek2 system (BioMerieux), respectively. Whole genome sequencing (WGS) was performed in an Illumina NextSeq2000 platform and MLST and resistome analysis of assembled genomes were performed by ResFinder, through the Center for Genomic Epidemiology platform. All isolates were resistant to ertapenem, imipenem, meropenem, and most to meropenem–varbobactam. Seventeen isolates belonged to the ST11 type and were positive for the OXA-48/NDM combination (by immunochromatography and NGS). Four isolates belonged to the ST39 type and were positive for the KPC/NDM combination (by immunochromatography and NGS). Finally, two isolates belonged to the ST258 type. One of them was positive for the OXA-48/KPC/NDM combination (by immunochromatography), but only bla_KPC was detected by WGS, and the second was positive for the OXA-48/KPC combination (by immunochromatography) and confirmed by WGS. This is the first report of an outbreak in Greece due to two simultaneous carbapenem-resistant populations harbouring double carbapenemases: a larger one comprising ST11 isolates harbouring the combination bla_NDM-1/bla_OXA-48, coupled by a smaller one comprising ST39 isolates harbouring the combination bla_KPC-2/bla_NDM-1. The implications of this particular situation regarding public health as well as intra-nosocomial infection prevention and control should be further monitored and studied. Full article

Carbapenem-resistant Gram-negative (CR-GN) pathogens pose a critical threat to patient outcomes in high-dependency and intensive care environments. This study aimed to delineate species prevalence, antimicrobial resistance phenotypes, carbapenemase genotypes, and clinical–environmental transmission dynamics across critical-care units. Cross-sectional surveillance was conducted in six ICUs and HDUs of a tertiary-care hospital in Karachi, Pakistan. We identified predominant species, quantified resistance patterns, and detected carbapenemase genes using PCR, exclusively on meropenem-resistant isolates. Network analysis highlighted high-centrality contamination hubs across ICUs and HDUs. Acinetobacter baumannii (36.7%) and Klebsiella pneumoniae (33.9%) were predominant, with 58% originating from environmental reservoirs. Meropenem non-susceptibility was 55% (60/109), and colistin non-susceptibility was 68.6% (35/51), based on standardized CLSI testing. ICU isolates exhibited significantly higher meropenem resistance than HDU isolates. Among carbapenem-resistant isolates, blaOXA-48-like (52.8%) and blaNDM (25%) were most prevalent. Network topology revealed ICU1 and HDU2 as high-centrality transmission nodes. These findings highlight pervasive environmental colonization and heightened antimicrobial pressure in ICUs, necessitating reinforced decontamination protocols, antimicrobial stewardship, and continuous molecular surveillance. This study provides the first integrated clinical–environmental surveillance of MDR Gram-negative bacteria in Pakistan, revealing that over half of isolates originated from surfaces and that network-based mapping can pinpoint contamination hubs driving hospital transmission. Full article

Carbapenem-resistant Klebsiella pneumoniae (CRKP) represents a critical global health threat, with ST101 identified as a major circulating clone in Saudi Arabia. We used whole genome sequencing and plasmid reconstruction to investigate the molecular characteristics of CRKP ST101 isolates from Saudi Arabia (2018–2021), analyzing antimicrobial resistance genes (ARGs), virulence factors, and plasmid structure and replicon types. Clinical isolates were obtained from the Ministry of National Guard Health Affairs (MNGHA) hospitals in Saudi Arabia between 2018 and 2021. Whole-genome sequencing was performed using the Illumina MiSeq^® platform, followed by comprehensive bioinformatic analysis of ARGs, virulence factors, and plasmid content. All ten isolates belonged to ST101 and harbored extensive antimicrobial resistance (AMR) and virulence determinants. Nine isolates (90%) carried bla_OXA-48, with three co-harboring bla_NDM-1, representing dual-carbapenemase producers. These carbapenemase genes were located on plasmids with distinct replicon types, including IncL/M, IncHI1B/IncFIB, and IncFIA/IncR. All isolates were multidrug-resistant (MDR), with half classified as extensively drug-resistant (XDR). Four isolates exhibited hypervirulent profiles, harboring aerobactin and yersiniabactin siderophores. This study provides comprehensive genomic characterization of CRKP ST101 in Saudi Arabia, revealing complex resistance mechanisms mediated by diverse plasmid types. The findings highlight the importance of genomic surveillance to track the evolution and dissemination of high-risk MDR and XDR lineages and inform targeted infection control strategies. Full article

Background: Rapid molecular detection of antimicrobial resistance (AMR) can shorten time to effective therapy in complicated urinary tract infections (cUTI), but the ability of gene presence and quantitative PCR signal (Ct, and ΔCt = Ct_marker − IC_Ct) to predict phenotypic non-susceptibility and clinical outcomes requires rigorous evaluation. We analyzed marker-level concordance, Ct→MIC relationships, and the clinical impact pathway in the randomized NCT06996301 trial. Methods: Marker–phenotype concordance metrics (sensitivity, specificity, PPV, NPV, LR+, LR−, κ) were computed for selected marker × species strata with stable sample sizes. Mixed-effects models (log₂[MIC] ~ ΔCt_marker + IC_Ct + collection_method + prior_abx + (1|site)) assessed quantitative Ct→MIC associations. ROC analyses evaluated ΔCt discrimination of phenotypic non-susceptibility. A pre-specified sensitivity analysis included smaller strata (n ≤ 20) with bootstrap 95% confidence intervals for ΔCt slopes and AUCs. Clinical analyses compared PCR-guided (n = 193) versus culture-guided (n = 169) arms for time-to-antibiotic and treatment success using adjusted logistic regression and causal mediation (time-to-antibiotic as mediator; bootstrap inference). Results: High genotype–phenotype concordance was observed for canonical markers (e.g., bla_CTX-M in E. coli: sensitivity 0.94 [95% CI 0.88–0.97], specificity 0.995 [95% CI 0.990–0.998], κ ≈ 0.93). Mixed models showed modest but significant Ct→MIC associations for select markers (e.g., bla_CTX-M in E. coli: ΔCt slope −0.15 [95% CI −0.27 to −0.02], p = 0.015). The sensitivity analysis (n ≤ 20 strata) confirmed consistent negative directions, with robust bootstrap CIs excluding zero for qnrS (E. coli), tetM (E. coli), blaNDM (Klebsiella), and qnrS (Proteus). ROC AUCs for ΔCt prediction of non-susceptibility ranged from 0.62 to 0.81 (95% CIs ≈ 0.47–0.97). Clinically, PCR guidance shortened median time to antibiotic initiation (20 h vs. 52 h) and increased treatment success (88.1% vs. 78.1%; adjusted OR 1.95 [95% CI 1.12–3.40], p = 0.018). Mediation analysis estimated that 63% (ACME 0.112 [95% CI 0.045–0.178], p = 0.002) of the treatment success benefit was mediated through earlier antibiotic initiation. Conclusions: Binary detection of high-impact AMR genes by multiplex PCR reliably predicts phenotypic non-susceptibility and accelerates effective therapy when integrated with stewardship workflows. Quantitative PCR (ΔCt) provides modest but reproducible information about MIC magnitude and may flag heteroresistant subpopulations. A pragmatic implementation model combining rapid PCR with conventional culture is recommended to optimize clinical benefit while retaining isolate recovery for definitive AST. Full article

Antimicrobial resistance in healthcare-associated infections represents one of the greatest threats to global health. The COVID-19 pandemic disrupted infection control and antimicrobial stewardship, potentially affecting the prevalence of pathogens and the development of resistance. This study aimed to investigate the prevalence, antimicrobial resistance, and clonal dissemination of ESKAPE pathogens isolated from bloodstream infections during the second year of the COVID-19 pandemic in four tertiary-care hospitals in Mexico. A total of 926 isolates were analyzed: Staphylococcus aureus (22.4%), Klebsiella pneumoniae (22%), Acinetobacter baumannii (21.5%), Pseudomonas aeruginosa (12.5%), Enterobacter cloacae (9.4%), Enterococcus faecalis (8.4%), and Enterococcus faecium (3.8%). High rates of multidrug resistance were observed in A. baumannii (70.9% XDR) and K. pneumoniae (71% XDR plus MDR with 79% ESBL). P. aeruginosa and E. cloacae showed the highest susceptibility rates (53% and 48%, respectively) to all antimicrobials. The main β-lactamases involved in resistance were bla_SHV, bla_CTX-M, and bla_TEM in K. pneumoniae, while the predominant carbapenemases were bla_OXA-24, bla_OXA-23 in A. baumannii, bla_NDM in K. pneumoniae, and bla_VIM in P. aeruginosa. Among Gram-positives, methicillin-resistant S. aureus accounted for 33.8% of isolates, and vancomycin resistance was higher in E. faecium (28%) than in E. faecalis (1.3%). Pulsed-field gel electrophoresis revealed endemic circulation of A. baumannii clones (Pulsotypes 1AC, 2AM), persistent for over a decade, and interhospital dissemination of S. aureus and K. pneumoniae clones. These findings underscore the epidemiological relevance of MDR ESKAPE pathogens during the COVID-19 pandemic and highlight the urgent need to optimize empirical therapy and maintain continuous genomic surveillance to enhance infection control in Mexican hospitals. Full article

This study developed a rapid and reliable SYBR-Green semiplex PCR assay for simulta-neous detection of major carbapenem resistance genes in Klebsiella pneumoniae and Acinetobacter baumannii. Two primer sets were used: one to detect blaKPC, blaNDM-1, and blaOXA-48 genes in K. pneumoniae and blaOXA-23 in A. baumannii, and another to amplify conserved 16S rRNA gene regions as internal controls. The intra- and inter-assay coeffi-cient of variation ranged from 0.03% to 3.8%. Standard curves exhibited excellent linearity across six logarithmic scales (10¹–10⁶ DNA copies/µL), with detection limits of 10–10² DNA copies/mL. Melting temperatures (Tm) were: 88.85 °C (KPIC), 90.65 °C (NDM-1), 89.45 °C (KPC), 84.23 °C (OXA-48), 87.81 °C (OXA-23), and 80.67 °C (ABIC). The SYBR-Green Semiplex PCR assay offers a rapid (65 min turnaround), cost-effective, and sensitive method for early detection of carbapenem-resistant pathogens, enabling timely targeted therapy and improved infection control by potentially reducing empirical antibiotic use before culture confirmation. Full article