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Applicability of Nanopore-only whole-genome sequencing for Pseudomonas aeruginosa outbreak investigation in the ICU setting: a multicentric study.

UNLABELLED: Pseudomonas aeruginosa outbreaks frequently occur in intensive care units (ICUs). In particular, ICU patients requiring mechanical ventilation are vulnerable to P. aeruginosa ventilator-associated pneumonia, which is associated with high morbidity and mortality. Fast and accurate genotyping during the early stage is crucial to document and manage P. aeruginosa outbreaks at the ICU. In this study, we have evaluated the applicability of Oxford Nanopore whole-genome sequencing (WGS) for outbreak investigation and antimicrobial resistance (AMR) prediction. To evaluate whether a Nanopore-only WGS workflow was able to reproduce Illumina-confirmed transmission clusters, 19 P. aeruginosa isolates from ICUs at UZ Brussels (Belgium) that were previously sequenced with Illumina were sequenced using a Nanopore-only workflow based on the latest V14 chemistry, followed by bioinformatic analysis via BugSeq and MBioSEQ Ridom Typer. Although both bioinformatic platforms showed high concordance between Illumina and Nanopore data, MBioSEQ Ridom Typer yielded the lowest allelic distance (maximum one cgMLST allele), confirming all outbreak clusters. When applying the Nanopore-only workflow to longitudinally collected isolates, low genetic heterogeneity (maximum three cgMLST alleles) was observed between isolates from the same patient. WGS and subsequent outbreak analysis of 65 respiratory P. aeruginosa isolates collected from 38 different ICU patients across six Belgian hospitals during a 9-month period showed no intra- or inter-hospital transmission. When the Nanopore-only WGS data were used to predict AMR, there was high categorical agreement (95%) between AMR genotype and phenotype. These findings highlight the potential of Nanopore WGS as a rapid and accurate tool for outbreak investigation of P. aeruginosa. IMPORTANCE: In recent years, Nanopore sequencing has found its way to clinical laboratories because of its affordability, scalability, and, most importantly, its ability to obtain sequencing results in near-real time. However, despite improved raw read accuracies with the latest generation R10.4.1 flow cells, the question remains whether the achieved accuracy is sufficient for accurate bacterial outbreak investigation, particularly in high-risk settings such as intensive care units (ICUs). In this study, we show that Nanopore-only whole-genome sequencing (WGS) is able to match Illumina-only WGS in terms of accuracy for Pseudomonas aeruginosa outbreak investigation in the ICU setting, although important sequence type-dependent and even strain-specific methylation issues need to be resolved in order to guarantee this accuracy. By providing a fast and accurate workflow for reliable P. aeruginosa outbreak investigation, this study could pave the way for large-scale implementation of Nanopore-only WGS, leading to faster outbreak response times.

Humans↗

Workflow modeling in critical care: piecing together your own puzzle.

The intensive care unit (ICU) is an instance of a very dynamic health care setting where critically ill patients are being managed. To provide good care, an extensive and coordinated communication amongst the role players, use of numerous information systems and operation of devices for monitoring and treatment purposes are required. The purpose of this research is to study error evolution and management within this environment. The focus is on representing the workflow of critical care environment, which emphasizes the importance such a representation may play in strategizing the management of medical errors. We used ethnographic observation and interview data to build individual pieces of the workflow, dependent on the individual and the activity concerned. Key personnel were intensively followed during their respective patient care activities and the related actions. All interactions were recorded for analysis. These clinicians and nurses were interviewed to complement the observation data and to delineate their individual workflows. These pieces of the ICU workflow were used to develop a generalize-able cognitive model to represent the intricate workflow applicable to other health care settings. The proposed model can be used to identify and characterize medical errors and for error prediction in practice.

Critical Care↗

Phage bioinformatics tools: a review of computational approaches for bacteriophage research.

Rising clinical interest in phage therapy and the exponential growth of metagenomic sequence catalogues have driven a rapid expansion of bacteriophage bioinformatics. More than 80 dedicated tools, mostly published since 2020, now span identification, assembly, annotation, taxonomy, lifestyle prediction, defence-system detection, and host prediction. Aimed at experienced practitioners and developers, this review synthesizes the field through the lens of three successive computational paradigms: sequence homology, bounded by database completeness; machine learning, constrained by labelled training data; and foundation models, which now achieve Matthews correlation coefficients above 0.95 in identification tasks and, through structure-informed prediction, raise functional annotation to over half of phage genes. Furthermore, we map the upstream components, namely, gene callers, homology engines, protein language models, and structural search tools, that underpin most downstream pipelines, exposing shared infrastructure and ecosystem-level fragility when dependencies change. To translate this into practice, we propose web-based and command-line reference workflows calibrated to user expertise and sample types. Finally, we set an agenda for the next wave of tool development. Roughly half of phage genes still resist functional annotation despite structural methods; no broadly generalizable strain-level host predictor exists for phage therapy; varying true-positive rates (0%-97%) underscore the absence of standardized community benchmarks analogous to Critical Assessment of Structure Prediction or Critical Assessment of Metagenome Interpretation. As generative genome models begin designing synthetic phages, progress will depend less on producing standalone tools than on rigorous evaluation, interoperable infrastructure, and clinically meaningful prediction targets.

Computational Biology↗

Integration of scanned document management with the anatomic pathology laboratory information system: analysis of benefits.

Electronic document management systems (EDMSs) have the potential to improve the efficiency of anatomic pathology laboratories. We implemented a novel but simple EDMS for scanned documents as part of our laboratory information system (AP-LIS) and collected cost-benefit data with the intention of discerning the value of such a system in general and whether integration with the AP-LIS is advantageous. We found that the direct financial benefits are modest but the indirect and intangible benefits are large. Benefits of time savings and access to data particularly accrued to pathologists and residents (3.8 h/d saved for 26 pathologists and residents). Integrating the scanned document management system (SDMS) into the AP-LIS has major advantages in terms of workflow and overall simplicity. This simple, integrated SDMS is an excellent value in a practice like ours, and many of the benefits likely apply in other practice settings.

Clinical Laboratory Information Systems↗

Picture archiving and communication system: the impact of filmless and distance radiology.

The progressive spread of Picture Archiving and Communication System (PACS) in medical imaging constitutes some of the major changes in the radiology and hospital environment during the past decade. The major benefit of PACS resides in its ability to communicate images and reports to referring physicians in a timely and reliable fashion. Filmless radiology offers the opportunity to redesign departmental and enterprise-wide workflow with increase in efficiencies of technologists, clerical staff and radiologists. PACS may improve patient care by providing real-time radiology and by enabling teleradiology. These technologies and the changing environment in radiology may force the radiologists to become more directly involved in the triage and management decisions for the patient. Occasional consultations, regularly scheduled consultation services, and multidisciplinary clinical discussions including a radiologist may help directing physicians to the best sequence of examinations to resolve a clinical problem.

Humans↗

Patient-controlled analgesic infusion pumps.

Patient-controlled analgesic (PCA) infusion pumps allow patients to self-administer doses of pain-relieving medication as needed, rather than having to summon a caregiver. The most significant risk when using these pumps is overmedication leading to narcotic-induced respiratory depression. Several current PCA pumps offer advanced error-reduction features designed to minimize the odds of an accidental overdose: dose error reduction systems, bar-code readers, and computer-based pump-programming systems that entail downloading infusion protocols via a hardwired connection to a PC. The availability of these features plays a dominant role in our ratings. We tested 11 pumps from seven suppliers. Two pumps are rated Preferred because they offer easy-to-use advanced error-reduction features that have been proven effective in a hospital setting. Two other pumps offer computer-based programming and are rated Acceptable, although we note that not all facilities may be able to accommodate the workflow necessary to use the computer-based programming. The remaining pumps are rated Not Recommended because they lack advanced error-reduction features; although most of these pumps perform acceptably, better choices are available.

Analgesia, Patient-Controlled↗

An application of digital network technology to medical image management.

With the advent of network technology, there is considerable interest within the medical community to manage the storage and distribution of medical images by digital means. Higher workflow efficiency leading to better patient care is one of the commonly cited outcomes [1,2]. However, due to the size of medical image files and the unique requirements in detail and resolution, medical image management poses special challenges. Storage requirements are usually large, which implies expenses or investment costs make digital networking projects financially out of reach for many clinical institutions. New advances in network technology and telecommunication, in conjunction with the decreasing cost in computer devices, have made digital image management achievable. In our institution, we have recently completed a pilot project to distribute medical images both within the physical confines of the clinical enterprise as well as outside the medical center campus. The design concept and the configuration of a comprehensive digital image network is described in this report.

Computer Communication Networks↗

Operational conditions affecting the vaccination of older adults.

BACKGROUND: The content and context of the process of vaccinating older adults against influenza in outpatient settings has not been adequately described. Failure to appreciate the causal antecedents or precursors to the act of provider recommendation may explain why so many efficacious interventions identified by the U.S. Task Force on Community Preventive Services fail to be routinely implemented and why influenza immunization rates have remained static over the past decade. METHODS: This study used critical path analysis from data collected during standardized workflow observations of patients more than 50 years of age from a convenience sample of 16 ambulatory care settings in San Diego, California; Rochester, New York; and Albuquerque, New Mexico. Observations were made from October 23, 2001 to January 31, 2002. RESULTS: In this study, 62% (151/243) of patients observed during scheduled extended visits received influenza vaccinations. When operational, temporal, and clinical factors are examined altogether through critical path analysis, a model of seven critical organizational support, temporal, and clinical activities emerges that is able to predict 93% of the immunizations. Variation from the model predicts 73% of the missed opportunities. CONCLUSIONS: Vaccination of adults should not be seen as simply an incremental activity added to the general health encounter. Assuring a high rate of vaccination requires adequate time and operational support. Provider-patient discussion is more productively viewed as the culmination of the immunization process, not the beginning. Finally, this study indicates the potential need to identify and compare processes of care associated with other specific preventive services.

Aged↗

Discharge communiqué: use of a workflow byproduct to generate an interim discharge summary.

Medical problems left unresolved during hospitalizations (along with recommended outpatient evaluations, test results pending at discharge, and discharge medication regimens) are often documented in patients' discharge summaries. However, studies have demonstrated that discharge summaries are frequently unavailable or inaccessible at post discharge visit(s). Interim discharge summaries have been shown to improve the flow of information between inpatient and outpatient physicians. We have constructed a web-based solution, discharge communiqués that are very much like interim discharge summaries but are an automatic byproduct of an every day workflow process, signout. The New SignOut System captures signout information and generates discharge communiqués immediately upon discharge. From June 2002-January 2003 7926 discharge communiqués were made available on 7926 patients and there were 12,920 look-ups of communiqués. Studies concur that 40-50% of patients will not have an available discharge summary making communiqués the primary source of clinical information on prior hospitalization for outpatient physicians.

Forms and Records Control↗

Evaluation of Cobas Integra 800 under simulated routine conditions in six laboratories.

The new selective access analyser Cobas Integra 800 from Roche Diagnostics was evaluated in an international multicentre study at six sites. Routine simulation experiments showed good performance and full functionality of the instrument and provocation of anomalous situations generated no problems. The new features on Cobas Integra 800, namely clot detection and dispensing control, worked according to specifications. The imprecision of Cobas Integra 800 fulfilled the proposed quality specifications regarding imprecision of analytical systems for clinical chemistry with few exceptions. Claims for linearity, drift, and carry-over were all within the defined specifications, except urea linearity. Interference exists in some cases, as could be expected due to the chemistries applied. Accuracy met the proposed quality specifications, except in some special cases. Method comparisons with Cobas Integra 700 showed good agreement; comparisons with other analysis systems yielded in several cases explicable deviations. Practicability of Cobas Integra 800 met or exceeded the requirements for more than 95% of all attributes rated. The strong points of the new analysis system were reagent handling, long stability of calibration curves, high number of tests on board, compatibility of the sample carrier to other Roche systems, and the sample integrity check for more reliable analytical results. The improvement of the workflow offered by the 5-position rack and STAT handling like on Cobas Integra 800 makes the instrument attractive for further consolidation in the medium-sized laboratory, for dedicated use of special analytes, and/or as back-up in the large routine laboratory.

Blood Proteins↗

Comparative performance of portable DNA extraction protocols and bioinformatics workflows for rapid detection of gram-negative bacteria and antimicrobial resistance using Oxford Nanopore sequencing.

Oxford Nanopore Technology (ONT) enables rapid, portable pathogen identification and antimicrobial resistance (AMR) detection, but the reliability of downstream genomic analyses is highly dependent on DNA extraction quality, particularly in resource-limited settings. This study comparatively evaluated four portable bacterial DNA extraction protocols derived from three commercial kits to determine their impact on nanopore sequencing performance, bioinformatics workflow completion, and field deployability. Six gram-negative bacterial isolates (Escherichia coli, n = 4; Pseudomonas sp., n = 1; and Salmonella sp., n = 1) were processed using four extraction protocols: SwiftX DNA, SwiftX DNA with proteinase K (ProtK), SwiftX ParaBact, and NucleoSpin Microbial. Twenty-four resulting DNA extracts were sequenced on a single multiplexed MinION R10.4.1 flow cell. Sequencing data were analyzed using validated Galaxy-based generic and species-specific pipelines. Workflow completion was defined as successful progression through quality control, assembly, virulence, plasmid, and AMR detection modules. DNA purity varied substantially by extraction protocol and was strongly associated with successful workflow completion (Kruskal-Wallis, P = 0.0006). Accordingly, NucleoSpin Microbial achieved 100% workflow completion, and SwiftX ParaBact achieved 83%, while both SwiftX DNA-based protocols failed to complete full workflows. Importantly, key AMR genes required to classify isolates as multidrug-resistant were consistently detected using both NucleoSpin Microbial and SwiftX ParaBact extractions. However, NucleoSpin Microbial assemblies showed significantly higher contiguity and enabled a broader, more complete detection of virulence factors, pathogenicity islands, plasmid replicons, and accessory AMR genes, reflecting enhanced genomic resolution.IMPORTANCERapid whole-genome sequencing is increasingly used to detect antimicrobial resistance and guide public health responses, but its reliability depends strongly on how bacterial DNA is extracted. In this study, we have shown that DNA extraction method choice has a major impact on Oxford Nanopore sequencing performance across clinically relevant gram-negative bacteria. While silica column-based extraction maximized genomic completeness and analytical depth, paramagnetic bead-based reverse purification offered superior portability with sufficient resolution for frontline AMR surveillance. These findings highlight a practical trade-off between field deployability and high-resolution genomic characterization in low-resource settings.

DNA extraction↗

Electronic patient records and the impact of the Internet.

The term electronic patient record (EPR) means the electronic collection of clinical narrative and diagnostic reports specific to an individual patient. A true EPR should allow physicians and nurses to practice in a paperless fashion. The wide adoption of Internet technologies should allow truly distributed sharing of patient data across traditional organizational barriers. Hence, the meaning of an EPR, as a representation of documents, should be transformed into a collaborative environment that supports workflow, enables new care models and allows secure access to distributed health data. This paper reviews the current realization of EPRs in the context of paper-based medical records. The Internet architecture that Boston-based medical informatics researchers refer to as W3-EMRS is described in the context of a successful implementation of CareWeb at the Beth Israel Deaconess Medical center. Finally, we describe how this Internet-based approach can be extended beyond the boundaries of traditional care settings to help evolve new collaborative models of eHealth.

Computer Security↗

Health information systems challenges: the Heidelberg conference and the future.

OBJECTIVES: To present a summary of the state of the art in Health Information Systems (HIS), as discussed during the Heidelberg HIS Working Group Conference, and to examine possible strategies for continuous improvement of the field. METHODS: The state of the art in HIS is briefly described, and the historical trends are examined that emerge from a review of the previous HIS Working Group conferences. To extrapolate from those trends and suggest possible new directions, we consider whether perceived difficulties in the diffusion of HIS systems are simply a product of technological factors, or whether fundamental social factors have been ignored. RESULTS: The experience with HIS environments is 'reasonably good' but not excellent, and true HIS success stories are not common. One of the apparent difficulties is that the typical HIS does not regard communication among clinical users as its core mission, even though repeated studies of information needs and practice patterns show that communication is the leading cost in today's health care environment. It is suggested that progress in the HIS arena will benefit from increased emphasis on the social aspects of health care, and better integration of diverse data to promote the organizational communication and workflow. Improvements could also come from a change to a highly participatory and evolutionary software engineering process, focusing on communication behavior and co-operative work practices. CONCLUSIONS: Future HIS development should view provider-provider and provider-patient communication as a core function, and incorporate features such as seamless support and tracking of communication, automatic gathering and presentation of data from multiple sources, data collection in atomic units by the most qualified provider, and integrated management, resource utilization, and tracking. The steps towards HIS that support these features might be smaller than generally suspected, once the appropriate changes in the reference point have been made.

Congresses as Topic↗

Good pharmacovigilance practices: technology enabled.

The assessment of spontaneous reports is most effective it is conducted within a defined and rigorous process. The framework for good pharmacovigilance process (GPVP) is proposed as a subset of good postmarketing surveillance process (GPMSP), a functional structure for both a public health and corporate risk management strategy. GPVP has good practices that implement each step within a defined process. These practices are designed to efficiently and effectively detect and alert the drug safety professional to new and potentially important information on drug-associated adverse reactions. These practices are enabled by applied technology designed specifically for the review and assessment of spontaneous reports. Specific practices include rules-based triage, active query prompts for severe organ insults, contextual single case evaluation, statistical proportionality and correlational checks, case-series analyses, and templates for signal work-up and interpretation. These practices and the overall GPVP are supported by state-of-the-art web-based systems with powerful analytical engines, workflow and audit trials to allow validated systems support for valid drug safety signalling efforts. It is also important to understand that a process has a defined set of steps and any one cannot stand independently. Specifically, advanced use of technical alerting methods in isolation can mislead and allow one to misunderstand priorities and relative value. In the end, pharmacovigilance is a clinical art and a component process to the science of pharmacoepidemiology and risk management.

Adverse Drug Reaction Reporting Systems↗

Rapid and accurate sepsis diagnostics via a novel probe-based multiplex real-time PCR system.

Sepsis is a critical clinical emergency that requires prompt diagnosis and intervention. Its prevalence has increased due to the aging population and increased antibiotic resistance. Early identification and the use of innovative technologies are crucial for improving patient outcomes. Modern methodologies are needed to minimize the turnaround time for diagnosis and improve outcomes. Rapid diagnostic tests and multiplex PCR are effective but have limitations in identifying a range of pathogens and target genes. Our study evaluated two novel probe-based multiplex real-time PCR systems: the SEPSI ID and SEPSI DR panels. These systems can quickly identify bacterial and fungal pathogens, alongside antibiotic resistance genes. The assays cover 29 microorganisms (gram-negative bacteria, gram-positive bacteria, yeast, and mold species), alongside 23 resistance genes and four virulence factors. A streamlined workflow uses 2 µL of broth from positive blood cultures (BCs) without nucleic acid extraction and provides results in approximately 1 h. We present the results from an evaluation of 228 BCs and 22 isolates previously characterized by whole-genome sequencing. In comparison to the reference methods, the SEPSI ID panel demonstrated a sensitivity of 96.88%, a specificity of 100%, and a PPV of 100%, whereas the SEPSI DR panel showed a sensitivity of 97.8%, a PPV of 89.7%, and a specificity of 96.7%. Both panels also identified additional pathogens and resistance-related targets not detected by conventional methods. This assay shows promise for rapidly and accurately diagnosing sepsis. Future studies should validate its performance in various clinical settings to enhance sepsis management and improve patient outcomes.IMPORTANCEWe present a new diagnostic method that enables the quick and precise identification of pathogens and resistance genes from positive blood cultures, eliminating the need for nucleic acid extraction. This technique can also be used on fresh pathogen cultures. It has the potential to greatly improve treatment protocols, leading to better patient outcomes, more responsible antibiotic use, and more efficient management of healthcare resources.

Humans↗

Electronic imaging and clinical implementation: work group approach at Mayo Clinic, Rochester.

Electronic imaging clinical implementation strategies and principles need to be developed as we move toward replacement of film-based radiology practices. During an 8-month period (1998 to 1999), an Electronic Imaging Clinical Implementation Work Group (EICIWG) was formed from sections of our department: Informatics Lab, Finance Committee, Management Section, Regional Practice Group, as well as several organ and image modality sections of the Department of Diagnostic Radiology. This group was formed to study and implement policies and strategies regarding implementation of electronic imaging into our practice. The following clinical practice issues were identified as key focus areas: (1) optimal electronic worklist organization; (2) how and when to link images with reports; (3) how to redistribute technical and professional relative value units (RVU); (4) how to facilitate future practice changes within our department regarding physical location and work redistribution; and (5) how to integrate off-campus imaging into on-campus workflow. The EICIWG divided their efforts into two phases. Phase I consisted of Fact finding and review of current practice patterns and current economic models, as well as radiology consulting needs. Phase II involved the development of recommendations, policies, and strategies for reengineering the radiology department to maintain current practice goals and use electronic imaging to improve practice patterns. The EICIWG concluded that electronic images should only be released with a formal report, except in emergent situations. Electronic worklists should support and maintain the physical presence of radiologists in critical areas and direct imaging to targeted subspecialists when possible. Case tools should be developed and used in radiology and hospital information systems (RIS/HIS) to monitor a number of parameters, including professional and technical RVU data. As communication standards improve, proper staffing models must be developed to facilitate electronic on-campus and off-campus consultation.

Diagnostic Imaging↗

[Teleradiology according to the x-ray ordinance--exemplary application of the open source software SecTelMed].

PURPOSE: Establishing a teleradiology connection according to the Rontgenverordnung (RoV) between a peripheral hospital and a university clinic will contribute to improve the radiological emergency treatment on weekends, holidays and at night. It will also better the medical care for patients and help to lower the costs. MATERIALS AND METHODS: By using the open source software SecTelMed, the radiological picture is transferred via an internet-linked SSH communication server. The integration of the teleradiological data transfer into the actual workflow is attained by using the already existing PACS infrastructure. While its installation is still under construction, it is actually employed for transferring a patient's preliminary investigations and obtaining second opinions. RESULTS: The project proves a reliable and affordable teleradiology according with RoV by using open source software. The applied infrastructure ensures an unproblematical teleradiological treatment as well as a safe picture transfer for medical consulting (second opinion) even for large CT examinations of 1,000 pictures and more. CONCLUSION: The implementation of the technical requirements on teleradiology according with the Rontgenverordnung can be reliably attained by applying the open source software SecTelMed.

Computer Communication Networks↗

Core genome and whole genome multi-locus sequence typing of Cronobacter isolates.

UNLABELLED: Cronobacter species, especially C. sakazakii and C. malonaticus, are opportunistic pathogens that are linked to severe infections in infants with high case fatality rates. In this study, we investigated whole genome sequencing (WGS) analysis approaches, specifically 7-gene multi-locus sequence typing (7-gene MLST), core genome MLST (cgMLST), and whole genome MLST (wgMLST) to subtype Cronobacter isolates. We analyzed a comprehensive set of 743 Cronobacter isolates derived from clinical, food, and environmental sources. We also evaluated high-quality single nucleotide polymorphism (hqSNP), cgMLST, and wgMLST to cluster epidemiologically related and differentiate sporadic C. sakazakii isolates. Our results indicate that both cgMLST and wgMLST accurately identify closely related isolates and are consistent with epidemiological findings. The allele-based analyses were also comparable with hqSNP analyses, the current gold standard. Our workflow also outputs 7-gene MLST allele calls, Cronobacter sequence types, and clonal complexes, which may be useful for historic comparisons during outbreak investigations. Following the recent classification of Cronobacter infections as nationally notifiable in the United States, our findings demonstrate the efficacy of WGS-based approaches within the PulseNet framework to improve outbreak detection and response strategies for Cronobacter. IMPORTANCE: Cronobacter species, specifically C. sakazakii and C. malonaticus, are opportunistic pathogens linked to severe infections in infants with high case fatality rates. This study highlights the critical importance of advanced molecular techniques in public health surveillance, using whole genome sequencing (WGS) methodologies such as multi-locus sequence typing (7-gene MLST), core genome MLST (cgMLST), and whole genome MLST (wgMLST). The validation of these WGS-based approaches within the PulseNet framework is timely, especially following the recent classification of Cronobacter infections as nationally notifiable in the United States. WGS methods not only enhance outbreak detection but can also inform public health guidance aimed at preventing infections and reducing mortality in vulnerable populations, especially infants. Our research supports implementation of cgMLST as a standardized approach for routine PulseNet surveillance of Cronobacter, with wgMLST and hqSNP analyses providing additional discriminatory power for outbreak investigations and high resolution phylogenetic analysis.

Multilocus Sequence Typing↗