Short Turnaround Time Research Articles

Association between blood culture turnaround time and clinical prognosis in emergency department patients with community acquired bloodstream infection: A retrospective study based on electronic medical records

ImportancePrevious investigations have found that time to positive blood culture (TTP) is a prognostic factor for clinical outcomes. In fact, what the emergency physician sees from the medical information system is TAT (turnaround time) defined as time required to post a bacterial culture report. We propose a definition of blood culture TAT that more closely aligns with clinical considerations by measuring the time from starting specimen culture to the release of an official blood culture report.We were curious to know whether the duration of TAT is as intricately linked to the prognosis of bacteremia as TTP. ObjectivesTo examine the association between TAT and outcomes of adult patients who present to the ED with community acquired bacteremia. DesignSetting, and Participants: This retrospective study utilized electronic medical records from Kaohsiung Veterans General Hospital (KVGH), a 1000-bed tertiary medical center in Taiwan. Patients were adults aged 18 years and older who presented to ED (Emergency department) for initial diagnosis of community acquired bacteremia from January 1, 2016 to March 31, 2021. Data analysis was performed from December 2022 to January 2023.Main outcomes and measures.The primary outcomes included mortality in the ED, all-cause in-hospital mortality, length of hospital stay, and all-cause 30-day mortality in relation to the individual first report of positive blood culture TAT. ResultsA total of 4011 eligible patients with bacteremia were evaluated, of which 207 patients had a blood culture TAT of ≤48 h. The overall 30-day all-cause mortality rate was 13%. Contrary to expectation, no statistically significant differences were observed in clinical prognosis between the TAT groups (≤48 versus >48 h). Subgroup analyses indicated that the length of TAT did not have a significant effect on clinical prognosis in patients who underwent lactate level assessment. Furthermore, no difference in clinical outcome was noted between TAT groups (≤48 versus >48 h) in terms of Gram-negative bacilli or Gram-positive cocci bacteremia. However, in patients with delayed antibiotic treatment (>3 h), a shorter TAT was significantly associated with a fatal outcome. ConclusionIn adults with community-acquired bacteremia, this study did not observe a significant association between blood culture TAT and clinical prognosis, except in cases of delayed antibiotic treatment.

Analyzing Breakthrough Infections in a Tertiary Care Hospital: A Retrospective Study Utilizing RAT-Based Screening

Prompt diagnosis of coronavirus disease 2019 (COVID-19) is requisite due to the upsurge of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) cases globally. Real-time reverse-transcription polymerase chain reaction (rRT-PCR) remains as the gold standard for the diagnosis of SARS-CoV-2 due to its higher sensitivity and specificity. Regardless, due to its longer turnaround time and expertise, bed side testing rapid antigen based tests were developed in order for portable, rapid diagnosis of the disease. The present study delineates the utility of Rapid Antigen Testing (RAT) screening among symptomatic and asymptomatic patients in a tertiary care hospital. This is a retrospective data collection study from Indian Council of Medical Research (ICMR) portal by exporting the excel sheet from March 2021 to March 2022 who undergone COVID testing in a tertiary care diagnostics facility. The demographic and vaccination details of the individuals were extracted using ICMR Sample referral form (SRF) from all patients who are tested for RAT. Of the total 9898 patients tested for RAT, 392 and 9506 were vaccinated and unvaccinated, respectively. Among them, number of positives was 3% and 15% respectively. The odds ratio showed that the vaccinated individuals had 81.4% reduced risk of getting infection compared to unvaccinated individuals (OR-0.186; p=0.001). A significant difference was observed between the type of vaccine used and COVID infection, majority of them had taken Covishield (83.5%) followed by Covaxin (16%) [p=0.0005]. Males showed a marginally significant p trend with COVID infectivity (OR=1.114, p=0.073) too. RAT is useful as a screening test for rapid diagnosis of infection in patients who are more prone to spread the disease. As RAT is cost effective and had short-turnaround time, it can be recommended in areas with upsurge in SARS-CoV-2 cases to administer isolation and infection control measures.

A Robust Closed-Tube Method for Resolving tp53M214K Genotypes.

The tp53M214K zebrafish mutant is a versatile platform with which to model a diverse spectrum of human diseases. However, currently available genotyping methods for this mutant require lengthy hands-on processes such as restriction digests and outsourced Sanger sequencing. To address this deficiency, we leveraged high-resolution melting analysis technology in conjunction with a parallel, in-tandem wild-type spike-in approach to develop a robust genotyping protocol capable of discriminating tp53M214K zygosity. In this study, we describe our method in detail. We anticipate that our genotyping protocol will benefit researchers utilizing the tp53M214K zebrafish mutant by offering reliable results with a shorter turnaround time, lower personnel involvement, and higher throughput than traditional methods, thereby decreasing the burden of genotyping and maximizing research efficiency.

CercaTest Red, a novel urine-based point-of-care test for the detection of preeclampsia

Objectives Preeclampsia (PE) poses a serious threat to the health of the pregnant woman and her developing fetus due to the difficulty in diagnosing the condition. The disease can develop and worsen suddenly without noticeable signs and symptoms. Thus, there is an urgent need for a simple Point of Care Test (POCT) that improves accessibility to testing and can be used as an aid in the diagnosis of PE. CercaTest Red is a noninvasive detection kit for impending preeclampsia using urine from pregnant women. This is especially pertinent for women who have limited access to secondary/tertiary healthcare as those in remote settings, low-income countries or simply lack of out of hours laboratory services. Methods The kit employs an absorptive column that separates Congo red dye bound to urinary misfolded protein from pregnant women and unbound dye. When a solution of Congo red dye pre-mixed with urine is loaded onto the absorptive matrix in a detection cuvette, the presence (positive) or absence (negative) of misfolded proteins can be determined based on the color of eluate collected in the lower section of the cuvette. 190 and 937 pregnant women who were >18 years old at the gestational age of ≥20 weeks were enrolled for the feasibility and validation cohort, respectively. The consistency between CercaTest Red and clinical diagnosis of PE according to the American College of Obstetricians and Gynecologist (ACOG) Guidelines was analyzed using the kappa statistic. Results The POCT has a limit of detection (LoD) of human urinary misfolded proteins equivalent to 0.45 μM of denatured human serum albumin, with high reproducibility and stability. An accuracy of 96.84% for diagnosis of preeclampsia with a Kappa statistic of 0.746 (p < 0.001) was validated in a cohort of 937 subjects. Conclusion This test is easy to use, cost-effective and portable with short turnaround time and no laboratory instrument requirement. In the future, the test may have the potential to become quantitative using spectroscopy (Chinese Clinical Trial Registry No. ChiCTR1800017692).

A genomic biomarker for the rapid identification of the rob(1;29) translocation in beef cattle breeds.

Robertsonian translocations, specifically rob(1;29) translocation, have reportedly been the most prevalent chromosomal abnormalities in cattle, affecting various breeds and leading to a decrease in fertility and reproductive value. Currently, the identification of rob(1;29) carriers relies on cytogenetic analysis that has limitations in terms of accessibility, cost, and sample requirements. To address these limitations, a novel genomic biomarker was developed in this study for the rapid and precise identification of rob(1;29) carriers. Using q-PCR, a specific copy number variation associated with translocation was targeted, which effectively distinguished between wild-type, homozygous and heterozygous carriers. Crucially, the biomarker can be applied to DNA extracted from various biological matrices, such as semen, embryos, oocytes, milk, saliva, coat, and muscle, and it is compatible with fresh, refrigerated, or frozen samples. Furthermore, this approach offers significant reductions in cost compared to those associated with traditional cytogenetic analysis and provides results within a short turnaround time. The successful development of this genomic biomarker has considerable potential for widespread adoption in screening programs. It facilitates timely identification and management of rob(1;29) carriers while mitigating economic losses and preserving genetic integrity in bovine populations.

Culture-independent identification of bloodstream infections from whole blood: prospective evaluation in specimens of known infection status.

Blood culture remains the "gold standard" for the diagnosis of sepsis/bloodstream infection (BSI) but has many limitations which may lead to a delay in appropriate and accurate treatment in patients. Molecular diagnostic methods have the potential for markedly improving the management of such patients through faster turnaround times and increased accuracy. But molecular diagnostic methods have not been widely adopted for the identification of BSIs. By incorporating a precursor step of selective lysis of host and non-viable microorganisms, our resistance and pathogen ID (RaPID)/BSI molecular assay addresses many limitations of blood culture and other molecular assay. The RaPID/BSI assay has an approximate turnaround time of 4 hours, thereby significantly reducing the time to appropriate and accurate diagnosis of causative microorganisms in such patients. The short turnaround time also allows for close to real-time tracking of pathogenic clearance of microorganisms from the blood of these patients or if a change of antimicrobial regimen is required. Thus, the RaPID/BSI molecular assay helps with optimization of antimicrobial stewardship; prompt and accurate diagnosis of sepsis/BSI could help target timely treatment and reduce mortality and morbidity in such patients.

Whole Blood Cardiac Troponin "Triaging" to Improve Early Detection of Myocardial Injury at a Pediatric Hospital.

The importance of offering on-site cardiac troponin (cTn) testing at pediatric hospitals may be underappreciated. We developed a rapid rule-in process for myocardial injury at a pediatric hospital experiencing delays in off-site high-sensitivity cardiac troponin T (hs-cTnT) testing. Collect-to-verify turnaround times (TATs) for off-site testing were reviewed. Pre-analytic changes to improve TATs were devised, implemented and evaluated, after which a new analyzer was selected and evaluated for on-site cTn testing. Performance of the new analyzer's assay was compared to the off-site hs-cTnT assay, and post go-live TATs for on-site testing were assessed. Median collect-to-verify TAT for short turnaround-time (STAT) priority off-site plasma hs-cTnT testing was 104 min, with 35% of orders having a TAT >120 min. Eliminating serum separator tubes and requiring a separate plasma separator tube did not significantly reduce TATs. A QuidelOrtho Triage® MeterPro whole blood cardiac troponin I (cTnI) assay was implemented to "triage" time-critical and STAT priority specimens collected for off-site hs-cTnT testing. Elevated cTnI (≥0.02 µg/L) had a sensitivity of 91% for clear elevations in hs-cTnT (≥53 ng/L) but a 0% sensitivity for modest elevations (5 to 13 ng/L, 14 to 52 ng/L). An interpretive comment was auto-appended to cTnI results indicating that clinicians should wait for the hs-cTnT result if cTnI was normal. Median collect-to-verify TAT for on-site cTnI testing was <50% the TAT for off-site hs-cTnT testing. On-site point-of-care whole blood cTn testing can rapidly confirm significant or late-presenting myocardial injury. Combined with simultaneous off-site high-sensitivity cardiac troponin (hs-cTn) testing, this workflow is a viable interim solution for pediatric hospitals without on-site hs-cTn testing.

New approaches to determine the technological challenged of oil fields at the late stage of development

Background: The main problem of the process of field development in the later stages is the limitation of free funds associated with the decline in production and the corresponding decrease in the profitable part of enterprises. At the same time, the accumulated effect of critical production problems is manifested, for the solution of which in practice a significant amount of capital investments is required in different areas of field development.&#x0D; The late stage of development of fields is associated with a number of problems, the main of which are the deterioration of the structure of the reserves and the slow pace of their replenishment, low withdrawal rates and high water cut, frequent failure of ground equipment, technological limitations of the production infrastructure, the quality of construction and completion of wells, a short turnaround time of production wells affecting the wells’ operation coefficient, and insufficient coverage of production processes with digitalization.&#x0D; Aim: Considering the large amount of work involved in the oil production process (from geology to onshore infrastructure), an internal benchmarking analytical tool is needed to identify current and most common technological challenges, to focus scientific, technical and production personnel in a certain direction and develop a systematic approach to solving production problems. The purpose of this work is to develop such a tool&#x0D; Materials and methods: As the input data, historical data on the production, hydrocarbons reserves, the current technological parameters of underground and surface equipment of 12 oil fields of the group of the NK “Kazmunaygaz” JSC companies were used. 17 basic criteria were formed in the areas of geology, development, oil production, drilling and infrastructure, which served as the basis for internal benchmarking of oil fields.&#x0D; Results: As a result of this work, a tool was developed for the diagnosis of key production problems of KMG oil fields. The diagnostic chart is effective for determining zones of the spread of production problems both in one oil field and in all oil fields of KMG. This approach can be scaled to the level of oil fields and horizons.&#x0D; Conclusion: The developed internal benchmarking tool serves as the basis for the annual analysis of key problems of the oil fields of the group of the NC “KazMunaygaz” JSC companies and the formation of a long -term plan to solve key production challenges.

A MALDI-TOF mass spectrometry-based method for detection of copy number variations in BRCA1 and BRCA2 genes.

Background: Identifying germline mutations in BRCA1 and BRCA2 genes (BRCAs) would benefit the carriers in multiple aspects. In addition to single-nucleotide variations and small indels, copy number variations (CNVs) is also an indispensable component of identifiable mutations in BRCAs. A sensitive, rapid and throughput-flexible method to detect CNVs would be preferred to meet the rising clinical requirements for BRCAs testing. Methods: We developed a MALDI-TOF-MS-based method (MS assay) which included three steps: first, multiplex end-point PCR followed by a single base extension reaction; second, automated analyte transfer and data acquisition; third, data analysis. We applied MS assay to detect CNVs in BRCAs in 293 Chinese patients with ovarian or pancreatic cancer. All the samples were examined by targeted next-generation sequencing (TS) simultaneously. Samples were further cross-validated by multiplex ligation-dependent probe amplification (MLPA) if the results from MS assay and TS were inconsistent. Long range PCR was then applied to identify the exact breakpoints in BRCAs. Results: MS assay introduced highly multiplexed panels to detect CNVs of BRCAs semi-quantitatively. Simplified on-board data analysis was available for MS assay and no complex bioinformatics was needed. The turnaround time of MS assay was less than 8hours with a hands-on time of only 40min. Compared to TS, MS assay exhibited higher sensitivity (100% vs. 75%) and was more flexible in throughput, with the reagent cost per sample remaining constant no matter how many samples were examined per assay. A total of eight CNVs in BRCAs were detected from the 293 samples, and the molecular breakpoints were successfully identified in five samples through long-range PCR followed by Sanger sequencing. Conclusion: Our results suggested that MS assay might be an effective method in primary screening for CNVs in genes such as BRCAs, especially when short turnaround time and/or high sensitivity is a top priority.

Optimization of the STARlet workflow for semi-automatic SARS-CoV-2 screening of swabs and deep respiratory materials using the RealAccurate Quadruplex SARS-CoV-2 PCR kit and Allplex SARS-CoV-2 PCR kit.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic triggered the implementation of large-scale screenings in the health care and in the general population. Consequently, medical laboratories have to apply lean laboratory management to design workflows that are able to process large batches within short turnaround times while maintaining flexibility to use different SARS-CoV-2 reverse transcription polymerase chain reactions (RT-PCRs) and to be able to process a variety of clinical samples. We validated two SARS-CoV-2 PCR assays on the STARlet workflow: Allplex SARS-CoV-2 PCR kit and RealAccurate Quadruplex SARS-CoV-2 PCR kit. Furthermore, we optimized and validated the STARlet workflow for semi-automatic screening for SARS-CoV-2 in upper respiratory swabs and deep respiratory materials (sputa, bronchoalveolar lavage, and aspirate). Strikingly, guanidine-containing lysis buffers allow for easy processing and can enhance sensitivity of SARS-COV-2 screening since sampling in these buffers may preserve viral transcripts as evident by the higher copy numbers of the SARS-CoV-2 N gene. Moreover, using the principles of lean laboratory management, several bottlenecks that are typical for medical laboratories were addressed. We show that lean laboratory management resulted in significant reduction of the turnaround times of the SARS-CoV-2 PCR in our laboratory. This report thus describes a useful framework for laboratories to implement similar semi-automated workflows.IMPORTANCEThe SARS-CoV-2 pandemic triggered the implementation of large-scale screenings in the health care and in the general population. Consequently, medical laboratories had to adapt and evolve workflows that are able to process large batches within short turnaround times while maintaining flexibility to use different assays and to be able to process a variety of clinical samples. We describe how the need for increased outputs and greater flexibility was addressed with respect to clinical samples and assays (Allplex SARS-CoV-2 PCR and RealAccurate Quadruplex SARS-CoV-2 PCR). Strikingly, we found that upper respiratory swabs collected in guanidine-containing lysis buffers both improved the ease of processing as well as enhanced the sensitivity of the SARS-CoV-2 screening. This report thus describes a useful framework for laboratories to implement and optimize similar semi-automated workflows.

End-point rapid detection of total and pathogenic Vibrio parahaemolyticus (tdh+ and/or trh1+ and/or trh2+) in raw seafood using a colorimetric loop-mediated isothermal amplification-xylenol orange technique.

Vibrio parahaemolyticus is the leading cause of bacterial seafood-borne gastroenteritis in humans worldwide. To ensure seafood safety and to minimize the occurrence of seafood-borne diseases, early detection of total V. parahaemolyticus (pathogenic and non-pathogenic strains) and pathogenic V. parahaemolyticus (tdh+ and/or trh1+ and/or trh2+) is required. This study further improved a loop-mediated isothermal amplification (LAMP) assay using xylenol orange (XO), a pH sensitive dye, to transform conventional LAMP into a one-step colorimetric assay giving visible results to the naked eye. LAMP-XO targeted rpoD for species specificity and tdh, trh1, and trh2 for pathogenic strains. Multiple hybrid inner primers (MHP) of LAMP primers for rpoD detection to complement the main primer set previously reported were designed by our group to maximize sensitivity and speed. Following the standard LAMP protocol, LAMP reaction temperature for rpoD, tdh, trh1, and trh2 detection was first determined using a turbidimeter. The acquired optimal temperature was subjected to optimize six parameters including dNTP mix, betaine, MgSO4, Bst 2.0 WarmStart DNA polymerase, reaction time and XO dye. The last parameter was done using a heat block. The color change of the LAMP-XO result from purple (negative) to yellow (positive) was monitored visually. The detection limits (DLs) of LAMP-XO using a 10-fold serial dilution of gDNA and spiked seafood samples were determined and compared with standard LAMP, PCR, and quantitative PCR (qPCR) assays. Subsequently, the LAMP-XO assay was validated with 102 raw seafood samples and the results were compared with PCR and qPCR assays. Under optimal conditions (65°C for 75 min), rpoD-LAMP-XO and tdh-LAMP-XO showed detection sensitivity at 102 copies of gDNA/reaction, or 10 folds greater than trh1-LAMP-XO and trh2-LAMP-XO. This level of sensitivity was similar to that of standard LAMP, comparable to that of the gold standard qPCR, and 10-100 times higher than that of PCR. In spiked samples, rpoD-LAMP-XO, tdh-LAMP-XO, and trh2-LAMP-XO could detect V. parahaemolyticus at 1 CFU/2.5 g spiked shrimp. Of 102 seafood samples, LAMP-XO was significantly more sensitive than PCR (P < 0.05) for tdh and trh2 detection and not significantly different from qPCR for all genes determined. The reliability of tdh-LAMP-XO and trh2-LAMP-XO to detect pathogenic V. parahaemolyticus was at 94.4% and 100%, respectively. To detect total and pathogenic V. parahaemolyticus, at least rpoD-LAMP-XO and trh2-LAMP-XO should be used, as both showed 100% sensitivity, specificity, and accuracy. With short turnaround time, ease, and reliability, LAMP-XO serves as a better alternative to PCR and qPCR for routine detection of V.parahaemolyticus in seafood. The concept of using a one-step LAMP-XO and MHP-LAMP to enhance efficiency of diagnostic performance of LAMP-based assays can be generally applied for detecting any gene of interest.

Lead with speed: Recasting the forensic laboratory product line and service delivery model

Forensic laboratories face a multitude of challenges when striving to deliver services to the criminal justice system. While many of these issues change over time, one in particular seems to endure the test of time … the need for faster results. Law enforcement wants and needs quicker response times to access critical information required to investigate their cases. One answer to this persistent problem is evolving technology. Technology not only permits a much quicker response than forensic laboratories are currently delivering, it can open the door to solving previously unsolvable cases. Along with applying new technology, an evaluation of current forensic laboratory product lines, service delivery models, and mindset regarding the role of forensic science-based investigative leads (termed forensic leads) is warranted. Resources and strategic planning are needed to realize the full potential of evolving technologies and what forensic laboratories can do to provide actionable and timely forensic leads to our criminal justice partners as a normal course of action instead of as an exception.This proposal is to establish a permanent, designated Forensic Lead Program (FLP) that resides under the umbrella of an accredited forensic laboratory and is tasked with the development and release of forensic leads. The FLP involves a focused menu of services, defined personnel roles, strict protocols, short turnaround time, standardized expectations, and targeted training, combined with the sense of urgency needed for consistent delivery of timely and actionable forensic leads. A dedicated FLP will save time and money by providing critical information for more focused investigations. ‘Speed is the need’ … for quick identification of those that threaten public safety and for the equally quick elimination of those wrongfully accused.Programs at two large state forensic laboratories will demonstrate how these concepts could be implemented along with their learning experiences. A business case will also be included to demonstrate the cost benefit of the Forensic Lead Program for DNA (CODIS – Combined DNA Index System) and NIBIN (National Integrated Ballistic Information Network), however other section services are expected to see similar benefits. Improving the response time by one day saves $1677.75 per $1 spent [1]. The return on investment (ROI) for applying DNA to firearms evidence returns $47.88 per $1 spent, or an 4,788 % ROI. Applying NIBIN (National Integrated Ballistic Information Network) to firearms evidence to provide investigative leads is $502.19 per $1 spent, which is a 50,219 % ROI. Recasting the forensic laboratory product line and service delivery model to ‘Lead with Speed’ makes both economic and investigative sense.

Utilization of Teleradiology by Intensive Care Units: A Cohort Study.

Imaging is indispensable to the diagnostic and treatment process. By facilitating access to rapid timely image interpretation, teleradiology plays a prominent role in improving access, quality of critical care, and management of the patients in intensive care units (ICU). The aim of the study is to investigate the role of teleradiology in ICU patient care and management. In our study, a total of 22,081 studies of a cohort of 14,900 patients which had been transmitted from intensive care units of 80 hospitals located across the United States of America through a teleradiology reporting workflow, were interpreted by the American Board Certified Radiologists empanelled by a teleradiology service provider, located in India. Among all modalities, the highest percentage of studies performed were computed tomography scan (47%) followed by radiographs (37.22%). Out of 22,081 cases under the study, 16,582 cases were reported during nighttime with a mean turnaround time (TAT) of 46.66 minutes 95% CI (46.27-47.04) while 5,499 cases were reported during daytime with a mean TAT of 44.66 minutes 95% CI (45.40-43.92). Setting up teleradiology service connectivity with a teleradiology service provider located in India, providing high-quality diagnostic interpretations and lower turnaround time with the ICUs in the US hospitals reduces the interval to intervention time and leads to efficient patient care management. Moreover, it also provides time advantage for US hospitals when on-site radiologists at night are unable to provide immediate coverage. The ICU teleradiology service model designed in the study would greatly help overcome the shortfall of radiologists in the hospitals, provide better patient management and care by quality reporting in short turnaround time, not only during daytime but also in the night hours or on holidays when on-site radiologists are unable to provide immediate coverage. Rao P, Mathur N, Kalyanpur A. Utilization of Teleradiology by Intensive Care Units: A Cohort Study. Indian J Crit Care Med 2024;28(1):20-25.

Examining the efficiency of porcine gastric mucin-coated magnetic beads in extraction of noroviruses from frozen berries

Human norovirus is the leading cause of foodborne gastroenteritis worldwide. Due to the low infectious dose of noroviruses, sensitive methodologies are required to detect and characterize small numbers of viral particles that are found in contaminated foods. The ISO 15216 method, which is internationally recognized for detection of foodborne viruses from high-risk food commodities, is based on viral precipitation, followed by RNA extraction and identification of the viral genome by RT-PCR. Although the ISO 15216 method is efficient, it is time consuming and tedious, does not report on the viral infectivity, and is sensitive to the presence of RT-PCR inhibitors. Norovirus capture by the porcine gastric mucin conjugated magnetic beads (PGM-MB) was developed as an alternative virus recovery method. It relies on the integrity of the viral capsid being able to bind to PGM. PGM contains a variety of histo-blood group antigens (HBGAs) that act as norovirus receptors. Therefore, the PGM-MB method allows for extraction of noroviruses, with potentially intact viral capsids, from complex food matrices. The viral genome can then be released through heat-shock of the captured virus. For this reason, we performed a parallel comparison between the ISO 15216 method and the PGM-MB method in isolation and quantification of noroviruses from frozen raspberries. We have demonstrated that the efficiency of the PGM-MB method in extraction of murine norovirus (MNV) and human norovirus GII.4 from raspberries is equal or better than the ISO 15216 method, while the PGM-MB has fewer steps and shorter turnaround time. Moreover, the PGM-MB method is more efficient in removing the inhibitors prior to RT-PCR analysis.

Mass spectrometry in clinical laboratory: applications and future prospects

Innovations in laboratory medicine face additional challenges due to the unmet need for extremely precise methods of disease diagnosis. Developments in mass spectrometry-based disease biomarker identification are constantly expanding the field of clinical diagnosis. More mass spectrometry-based in vitro diagnostics are anticipated to move from the bench to the bedside shortly, although several are currently incorporated into standard clinical procedures. The molecular detection by mass spectrometry technique is very effective in diagnosis of the disease and therapeutic monitoring because of its extremely high sensitivity, specificity, and short turnaround time. Mass spectrometry is a potent analytical instrument that may be used to analyse a variety of materials and matrices; its use in clinical laboratory medicine applications is rising. For clinical prognostics and diagnostic purposes, mass spectrometry imaging has been widely utilized to distinguish between healthy and diseased tissues. Modern single-cell studies will benefit from its cutting-edge applications, which will offer comprehensive cellular biochemical data for mechanistic comprehension and, eventually, the development of therapeutic interventions. Within the field of laboratory medicine, the application of mass spectrometry is crucial, especially in terms of diagnostics and therapeutic drug monitoring. This review on the clinical application of mass spectrometry and its future prospects promises an insightful exploration of the evolving landscape of mass spectrometry in clinical settings, delving into its current applications and shedding light on its potential future advancements, for which a comprehensive search in the PubMed, ScienceDirect, and Web of Science databases was conducted.

E-HRM: Learning approaches, applications and the role of artificial intelligence

E-HRM (Electronic Human Resources Management), which is derived from the concept of HRM (Human Resources Management) plays a significant role in automating certain key processes in the department of human resources. One of the modules of E-HRM is the training or the learning module, which when combined with a digital source turns out to be an E-Learning (Electronic Learning) or E-Training (Electronic Training) module. This is a transformation of converting the learning platform from an offline to an online mode. The organizations to increase their level of training from their employees should look for a fast-paced solution at a shorter turn-around time and the prime way to perform such a strategy is to automate the whole process of training and predict the training need and outcome. This research paper is focused on two aspects of e-learning i.e., how an e-learning system is collaborated with an intelligent system in the form of Artificial Intelligence and the other aspect is how an employee turn over data fetched from organizations in the IT (Information Technology) sector can help understand the real requirements of learning among employees in the IT organizations.

(Invited) Nanoplasmonic Optofluidic Biosensors for Rapid and Precise Immune Functional Analysis in Personalized Immunotherapy

Cytokines, acting as intercellular mediators and modulators, are essential biomolecules that regulate diverse functions within the immune response. The rapid and accurate quantification of cytokine-based immune fingerprints is vital for effectively treating immune-related diseases, particularly in point-of-care settings where immediate treatment decisions depend on precise determinations of an individual patient's immune status. Addressing the pressing need for cytokine immunoassays that offer high sensitivity, throughput, multiplexing capability, and short turnaround times at low system complexity, this presentation will discuss novel plasmonic nanomaterial-based optofluidic cytokine biosensors for immune functional analysis ranging from whole blood to single-cell levels. The multiscale experimental and theoretical research will bridge the gap in fundamental understanding of the immune system, enhancing the applicability, diagnosis, and predictive power for immune diseases. The developed platforms aim to provide biologists and clinicians with the capability to real-time monitor patients' immune statuses, a transformative achievement with immense potential for safe, effective, and personalized immunotherapy.

Evaluation of the Aggressive-Variant Prostate Cancer Molecular Signature in Clinical Laboratory Improvement Amendments (CLIA) Environments.

Aggressive-variant prostate cancers (AVPCs) are a subset of metastatic castrate-resistant prostate cancers (mCRPCs) characterized by defects in ≥ two of three of TP53, RB1, and PTEN (AVPCm), a profile linked to lineage plasticity, androgen indifference, and platinum sensitivity. Men with mCRPC undergoing biopsies for progression were assessed for AVPCm using immunohistochemistry (IHC), next-generation sequencing (NGS) of solid tumor DNA (stDNA), and NGS of circulating tumor DNA (ctDNA) assays in CLIA-certified labs. Biopsy characteristics, turnaround times, inter-reader concordance, and inter-assay concordance were assessed. AVPCm was detected in 13 (27%) patients via IHC, two (6%) based on stDNA, and seven (39%) based on ctDNA. The concordance of the IHC reads between pathologists was variable. IHC had a higher detection rate of AVPCm+ tumors with the shortest turnaround times. stDNA had challenges with copy number loss detection, limiting its detection rate. ctDNA detected the greatest proportion of AVPCm+ tumors but had a low tumor content in two thirds of patients. These data show the operational characteristics of AVPCm detection using various assays, and inform trial design using AVPCm as a criterion for patient selection or stratification.

Clinical Evaluation of Direct Reverse Transcription PCR for Detection of SARS-CoV-2 Compared to Conventional RT-PCR in Patients with Positive Rapid Antigen Test Results during Circulation of Emerging Viral Variants.

The emergence of the Omicron (B.1.1.529) variant of SARS-CoV-2 has precipitated a new global wave of the COVID-19 pandemic. The rapid identification of SARS-CoV-2 infection is imperative for the effective mitigation of transmission. Diagnostic modalities such as rapid antigen testing and real-time reverse transcription polymerase chain reaction (RT-PCR) offer expedient turnaround times of 10-15 min and straightforward implementation. This preliminary study assessed the correlation between outcomes of commercially available rapid antigen tests for home use and conventional reverse transcription polymerase chain reaction (RT-PCR) assays using a limited set of clinical specimens. Patients aged 5-99 years presenting to the emergency department for SARS-CoV-2 testing were eligible for enrollment (n = 5652). Direct PCR and conventional RT-PCR were utilized for the detection of SARS-CoV-2. The entire cohort of 5652 clinical specimens was assessed by both modalities to determine the clinical utility of the direct RT-PCR assay. Timely confirmation of SARS-CoV-2 infection may attenuate viral propagation and guide therapeutic interventions. Additionally, direct RT-PCR as a secondary confirmatory test for at-home rapid antigen test results demonstrated sensitivity comparable to conventional RT-PCR, indicating utility for implementation in laboratories globally, especially in resource-limited settings with constraints on reagents, equipment, and skilled personnel. In summary, direct RT-PCR enables the detection of SARS-CoV-2 with a sensitivity approaching that of conventional RT-PCR while offering expedient throughput and shorter turnaround times. Moreover, direct RT-PCR provides an open-source option for diagnostic laboratories worldwide, particularly in low- and middle-income countries.

Rapid haemoglobin A and S quantification using Tosoh HLC-723G8 in variant mode for patients with sickle cell disease.

The management of life-threatening complications in patients with sickle cell disease (SCD) requires an accurate and reproducible quantification of haemoglobin A (HbA) and S (HbS) with a short turnaround time and 24-7 availability. We propose a novel method for quantifying HbA and HbS using the glycated haemoglobin (HbA1c) assay on a Tosoh HLC-723G8 (G8) analyser in variant mode. HbA and HbS results obtained using our method highly correlated with results obtained using a reference method (r > 0.99 for 124 samples of patients with SCD or sickle cell trait). Our method met laboratory requirements for linearity (coefficient of variation [CV] and bias <5%), between-run and within-run reproducibility (CV <10%) and carryover (<0.5%) over the range of HbS and HbA values expected in a therapeutic context. Using the G8 analyser in variant mode is viable for monitoring HbA and HbS concentrations in dire situations. This method is easy to use, quick (1.6min per sample), and automatable and produces highly reproducible results without significant bias. Finally, it does not require modifications to the analytical pipeline recommended by the supplier, enabling a 24-7 availability without disrupting routine monitoring of HbA1c in the laboratory.