Bacterial Infections and Antimicrobial Resistance: Diagnosis and Management

A special issue of Diagnostics (ISSN 2075-4418). This special issue belongs to the section "Diagnostic Microbiology and Infectious Disease".

Deadline for manuscript submissions: closed (30 November 2025) | Viewed by 3166

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Guest Editor
National Meningitis Reference Laboratory, Department of Public Health Policy, School of Public Health, University of West Attica, Athens, Greece
Interests: bacterial meningitis; N. meningitidis; S. pneumoniae; H. influenzae
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Special Issue Information

Dear Colleagues,

This Special Issue addresses the growing global challenge of bacterial infections and the alarming rise of antimicrobial resistance (AMR). It explores innovative diagnostic approaches, including rapid molecular techniques, next-generation sequencing, and point-of-care testing, which enable early and accurate identification of pathogens and resistance patterns. This Special Issue also highlights the critical role of antimicrobial stewardship programs in optimizing the use of antibiotics and combating resistance. Advances in alternative therapies, such as phage therapy, immunotherapies, and novel antimicrobial agents, are discussed as promising strategies to address multidrug-resistant infections. Additionally, the integration of artificial intelligence and machine learning in infection management are examined for their potential to enhance diagnostics and treatment decision-making. Contributions from leading experts emphasize the need for a multidisciplinary approach, combining microbiology, clinical practice, and public health initiatives, to tackle AMR effectively. This collection not only provides a comprehensive overview of current challenges but also outlines future directions for research and policy, aiming to improve patient outcomes and mitigate the global threat of AMR.

Prof. Dr. Georgina Tzanakaki
Guest Editor

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Keywords

  • bacterial infections
  • antimicrobial resistance (AMR)
  • diagnostic innovations
  • molecular diagnostics
  • antimicrobial stewardship
  • multidrug-resistant pathogens
  • alternative therapies
  • artificial intelligence

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Published Papers (4 papers)

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Research

14 pages, 230 KB  
Article
Comparison of Bacteriology Between Geriatric and Adult Rhinosinusitis with and Without NPs
by Meng-Chun Lin, Yi-Ching Chen and Rong-San Jiang
Diagnostics 2025, 15(23), 3035; https://doi.org/10.3390/diagnostics15233035 - 28 Nov 2025
Viewed by 163
Abstract
Background: In this study, we attempted to compare the bacteriology of chronic rhinosisusitis (CRS) with and without nasal polyps between geriatric and adult patients. Methods: This retrospective cross-sectional study included 751 patients with CRS who underwent bilateral primary functional endoscopic sinus [...] Read more.
Background: In this study, we attempted to compare the bacteriology of chronic rhinosisusitis (CRS) with and without nasal polyps between geriatric and adult patients. Methods: This retrospective cross-sectional study included 751 patients with CRS who underwent bilateral primary functional endoscopic sinus surgery. Before surgery, swab samples were collected from the middle meatus for bacterial cultures using cotton-tipped sticks. Subjects were divided into adult (20 to 64 years, n = 683) and elderly (65 years, n = 68) groups. The results of the bacteria culture were analyzed according to age group and the presence of nasal polyps. Results: The bacterial culture rate was higher in geriatric patients (55.9%) than in adults (44.9%), but the difference was not statistically significant. However, geriatric patients showed a higher bacterial culture rate (57.6%) than adult patients (29.6%) without nasal polyps. This difference was statistically significant. Conclusions: Geriatric patients with CRS exhibited higher bacterial culture rates, particularly on the non-polyp side. These findings suggest a possible age-related susceptibility to microbial colonization, underscoring the need for age-specific infection management strategies. Full article
24 pages, 2524 KB  
Article
Phenotype-First Diagnostic Framework for Tracking Fluoroquinolone Resistance in Escherichia coli
by Eman Marzouk and Abdulaziz M. Almuzaini
Diagnostics 2025, 15(22), 2831; https://doi.org/10.3390/diagnostics15222831 - 7 Nov 2025
Viewed by 476
Abstract
Background: Fluoroquinolone (FQ) resistance in Escherichia coli (E. coli) undermines empiric therapy and often coincides with multidrug resistance (MDR). Because sequencing is not routinely available in many laboratories, we evaluated a phenotype-first, sequencing-independent diagnostic framework deployable on standard platforms. Methods: We [...] Read more.
Background: Fluoroquinolone (FQ) resistance in Escherichia coli (E. coli) undermines empiric therapy and often coincides with multidrug resistance (MDR). Because sequencing is not routinely available in many laboratories, we evaluated a phenotype-first, sequencing-independent diagnostic framework deployable on standard platforms. Methods: We profiled 45 archived E. coli isolates for susceptibility (Clinical and Laboratory Standards Institute [CLSI]-guided), extended-spectrum β-lactamase (ESBL) and AmpC β-lactamase (AmpC) phenotypes, MDR, and multiple-antibiotic resistance (MAR) indices. Ten founders (five FQ-susceptible [FQ-S], five low-level resistant [LLR]) seeded 20 parallel lineages exposed to stepwise ciprofloxacin. We tracked minimum inhibitory concentrations (MICs), collateral resistance, growth kinetics, and biofilm biomass using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identification, automated and reference antimicrobial susceptibility testing (AST), growth-curve analysis, and crystal violet microtiter assays. The intended use is a sequencing-independent workflow for routine laboratories—especially where whole-genome sequencing is not readily available—working with archived or prospective clinical E. coli. This workflow is best applied when local FQ nonsusceptibility threatens empiric reliability; inputs include standard ID/AST with simple growth and biofilm assays. Primary outputs include: (i) MIC trajectories with time to high-level resistance (HLR), (ii) ΔMAR-summarized collateral resistance with class-level susceptible-to-resistant conversions, and (iii) concise fitness/biofilm summaries to guide empiric-policy refresh and early de-escalation. Results: At baseline, ciprofloxacin nonsusceptibility was 40.0%; ESBL and AmpC phenotypes were confirmed in 28.9% and 15.6%, respectively; 46.7% met the MDR definition; and the median MAR index was 0.29. During evolution, 70% of lineages reached HLR (MIC ≥ 4 μg/mL), with earlier conversion from LLR versus FQ-S founders (median 7 vs. 11 passages). Collateral resistance emerged most often to third-generation cephalosporins (3GCs), trimethoprim–sulfamethoxazole, and tetracyclines, while carbapenem activity was preserved. MAR increased in parallel with rising MICs. Resistance acquisition imposed modest fitness costs (slightly reduced growth rates and longer lag phases) that were partly offset under subinhibitory ciprofloxacin, whereas biofilm biomass changed little. Conclusions: this phenotype-first, routine-laboratory workflow rapidly maps FQ resistance and clinically relevant co-selection in E. coli. In high-resistance settings, empiric FQ use is difficult to justify, and MAR trends provide practical co-selection signals for stewardship. This reproducible framework complements genomic surveillance and is directly applicable where sequencing is unavailable. Full article
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12 pages, 1038 KB  
Article
Rapid Identification of Carbapenemase Genes Directly from Blood Culture Samples
by Ghada A. Ziad, Deena Jalal, Mohamed Hashem, Ahmed A. Sayed, Sally Mahfouz, Ahmed Bayoumi, Maryam Lotfi, Omneya Hassanain, May Tolba, Youssef Madney, Lobna Shalaby and Mervat Elanany
Diagnostics 2025, 15(19), 2480; https://doi.org/10.3390/diagnostics15192480 - 28 Sep 2025
Viewed by 936
Abstract
Background/Objectives: The rapid identification of carbapenemase genes directly from positive blood culture (BC) samples shortens the time needed to initiate optimal antimicrobial therapy for Carbapenemase-Producing Enterobacterales (CPE) infections. Several commercial automated PCR systems are available for detecting CPE resistance genes but are expensive. [...] Read more.
Background/Objectives: The rapid identification of carbapenemase genes directly from positive blood culture (BC) samples shortens the time needed to initiate optimal antimicrobial therapy for Carbapenemase-Producing Enterobacterales (CPE) infections. Several commercial automated PCR systems are available for detecting CPE resistance genes but are expensive. The Xpert® Carba-R assay (Cepheid GeneXpert System) has high sensitivity and specificity for the detection of carbapenamase genes from bacterial colonies or rectal swabs, with an affordable price. This assay was not used for positive BC testing of CPE resistance genes. Whole-Genome Sequencing (WGS) for resistance genes can be used as the gold standard at a research level. In this study, we evaluated the performance of the Xpert® Carba-R assay for the early detection of carbapenamase genes directly from positive BCs, using WGS as the gold standard. Methods: A prospective observational study was conducted at Children’s Cancer Hospital-Egypt (CCHE-57357). All positive BCs underwent direct gram staining and conventional cultures. A total of 590 positive BCs containing Gram-negative rods (GNRs) were identified. The Xpert® Carba-R assay was used to detect carbapenemase genes directly from the positive BC bottle compared with WGS results. Results: Among the 590 GNR specimens, 178 were found to carry carbapenemase genes using the Xpert® Carba-R assay, with results obtained in approximately one hour. The main genotypes detected were blaNDM, blaOXA-48-like, and dual blaNDM/blaOXA-48-like at 27%, 29%, and 33%, respectively. The agreement between Xpert® Carba-R assay and WGS results was almost perfect for the genotype resistance pattern of isolates and individual gene detection. Conclusions: The use of the Xpert® Carba-R assay directly from BC bottles was an easy-to-use, time-saving, affordable tool with high accuracy in identifying carbapenemase genes and, thus, shortens the time needed to initiate optimal antimicrobial therapy for CPE infections. Full article
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14 pages, 3168 KB  
Article
Development of SNP-LAMP Combined with Lateral Flow Dipstick to Detect the S531L rpoB Gene Mutation in Rifampicin-Resistant Mycobacterium tuberculosis
by Jutturong Ckumdee, Monpat Chamnanphom, Supaporn Wiwattanakul, Somchai Santiwatanakul, Kwanchai Onruang and Thongchai Kaewphinit
Diagnostics 2025, 15(17), 2183; https://doi.org/10.3390/diagnostics15172183 - 28 Aug 2025
Viewed by 1193
Abstract
Background: Tuberculosis (TB) remains a primary global health concern, despite the widespread availability of effective chemotherapeutic interventions. The emergence and dissemination of drug-resistant strains of Mycobacterium tuberculosis, particularly those exhibiting resistance to rifampicin, present significant obstacles to the success of TB control [...] Read more.
Background: Tuberculosis (TB) remains a primary global health concern, despite the widespread availability of effective chemotherapeutic interventions. The emergence and dissemination of drug-resistant strains of Mycobacterium tuberculosis, particularly those exhibiting resistance to rifampicin, present significant obstacles to the success of TB control programs. Consequently, there is an urgent need for rapid, sensitive, and specific molecular diagnostic tools to inform timely clinical decision-making and reduce the transmission of disease. Loop-mediated isothermal amplification (LAMP) has gained attention as a promising alternative to conventional polymerase chain reaction (PCR) techniques. This method, which facilitates DNA amplification under constant temperature conditions, offers advantages including high specificity, rapid turnaround time, and operational simplicity—features that render it especially suitable for implementation in resource-limited settings. Methods: In this study, a LAMP assay targeting the rpoB gene was developed, with particular focus on detecting the codon 531 C→T mutation associated with rifampicin resistance. A set of four to six primers was designed to recognize six distinct regions of the target sequence. Allele-specific amplification was achieved by incorporating a deliberate single nucleotide mismatch at the 3′ terminus of the B2 primer to enable precise discrimination between wild-type and mutant alleles. The assay was conducted at an optimized temperature of 61 °C for 60 min, followed by visual detection using a lateral flow dipstick (LFD) within five minutes. Results: The LAMP-LFD assay demonstrated 100% concordance with drug susceptibility testing (DST) and DNA sequencing. No cross-reactivity with wild-type strains was observed, underscoring the assay’s high specificity. Conclusions: This platform offers a robust, field-deployable solution for detecting the codon 531 C→T mutation associated with rifampicin resistance in low-resource settings. Full article
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