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Use of IR Biotyper as a feasible methodology to type Klebsiella pneumoniae.

UNLABELLED: Klebsiella pneumoniae is one of the most frequently reported healthcare-associated pathogens. The current gold standard approach to perform the epidemiological typing of these bacteria is Whole Genome Sequencing (WGS), which is an expensive and challenging procedure. IR Biotyper (Bruker Daltonics, GmbH) is a new equipment based on Fourier transform infrared spectroscopy, which allows a rapid, low-cost, and user-friendly method to type bacterial isolates. However, there is a need for studies that evaluate the efficacy of the IR Biotyper. The aim of this study was to evaluate the capability of IR Biotyper to type K. pneumoniae according to sequence type (ST) and capsular type-using K locus (KL)-as well as to develop a classifier using machine learning. Seventy-three isolates of K. pneumoniae previously characterized by WGS were selected for IR Biotyper analysis using principal component analysis for dimensionality reduction, Euclidean, and unweighted pair group method with arithmetic mean (UPGMA) for clustering method, and spectra were analyzed in the 1,300-800 cm⁻¹ wavenumber range. Among these, 54 isolates were used to create a classifier, and 19 were used to validate the classifier. When considering the ST, ST307 was grouped in the same cluster as ST11. When KL was considered for the analysis, the clusters were 100% correctly grouped according to their KL type. Furthermore, the classifier developed was able to classify the isolates according to KL with a high concordance. This study showed that KL correlates well with KL for typing K. pneumoniae isolates using the IR Biotyper. Additionally, IR Biotyper demonstrated to be a cost-effective method and a promising tool to classify isolates within minutes. IMPORTANCE: Klebsiella pneumoniae is a major cause of severe hospital infections, and controlling its spread requires quick identification and comparison of bacterial strains. WGS is accurate but expensive, slow, and technically demanding. In this study, we evaluated the IR Biotyper, a device that uses infrared light to analyze bacteria and group them by capsule type-a key feature linked to their spread. The IR Biotyper matched WGS results with high accuracy, delivering results in minutes instead of days. This fast, affordable method can help hospitals detect outbreaks earlier and respond more effectively. Our findings suggest that the IR Biotyper is a valuable tool for routine use in microbiology laboratories, supporting epidemiological surveillance and outbreak control.

Klebsiella pneumoniae

Performance of the IR Biotyper, Nanopore, and Illumina sequencing to discriminate Escherichia coli strains originating from poultry.

UNLABELLED: Escherichia coli is a highly diverse bacterial species that includes avian pathogenic E. coli (APEC), one of the most prevalent causative agents of disease in poultry worldwide. Rapid and accurate discrimination of E. coli strains is essential for outbreak management, antimicrobial resistance surveillance, and vaccine development. In this study, we compared the performance of Fourier Transform Infrared (FTIR) spectroscopy using the IR Biotyper system with Nanopore and Illumina whole-genome sequencing (WGS) for typing 200 E. coli isolates, originating from four poultry rearing farms in the Netherlands. From each farm, we sampled 10 one-day-old meat type rearing chicks, and from every chick, we isolated 5 E. coli strains. FTIR clustering showed strong concordance with WGS-based classifications, particularly serotyping and core-genome similarity determined by PopPUNK analysis (Adjusted Rand Index 0.75-0.92). While Nanopore and Illumina sequencing provided the highest genetic resolution, FTIR offered a faster (max 6 vs 12-28 days for 200 isolates) and more cost-effective alternative for assessing clonality. Across all methods, multiple strains were detected per farm, whereas most birds carried a single dominant E. coli strain. Our findings demonstrate that FTIR provides a reliable and scalable phenotypic method for rapid strain discrimination in E. coli, complementing WGS in diagnostic, surveillance, and epidemiological settings where speed and throughput are critical. IMPORTANCE: Escherichia coli is a major pathogen in poultry and a potential zoonotic risk for humans. Rapid and accurate discrimination of avian pathogenic E. coli (APEC) strains is critical for outbreak management, antimicrobial resistance surveillance, and the design of effective autogenous vaccines. In this study, we compared Fourier Transform Infrared (FTIR) spectroscopy with Nanopore and Illumina whole-genome sequencing for strain typing of E. coli isolates originating from poultry. The results show that FTIR provides comparable clustering accuracy to genomic approaches at a fraction of the time and costs. This work demonstrates that FTIR can serve as a practical, high-throughput alternative for routine monitoring of E. coli in veterinary diagnostics and food safety of poultry meat, enabling faster decision-making and more targeted interventions across the poultry production chain.

Animals