PATHPOD - A loop-mediated isothermal amplification (LAMP)-based point-of-care system for rapid clinical detection of SARS-CoV-2 in hospitals in Denmark.

Sensitive and rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a vital goal in the ongoing COVID-19 pandemic. We present in this comprehensive work, for the first time, detailed fabrication and clinical validation of a point of care (PoC) device for rapid, onsite detection of SARS-CoV-2 using a real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) reaction on a polymer cartridge. The PoC system, namely PATHPOD, consisting of a standalone device (weight less than 1.2 kg) and a cartridge, can perform the detection of 10 different samples and two controls in less than 50 min, which is much more rapid than the golden standard real-time reverse-transcription Polymerase Chain Reaction (RT-PCR), typically taking 16-48 h. The novel total internal reflection (TIR) scheme and the reactions inside the cartridge in the PoC device allow monitoring of the diagnostic results in real-time and onsite. The analytical sensitivity and specificity of the PoC test are comparable with the current RT-PCR, with a limit of detection (LOD) down to 30-50 viral genome copies. The robustness of the PATHPOD PoC system has been confirmed by analyzing 398 clinical samples initially examined in two hospitals in Denmark. The clinical sensitivity and specificity of these tests are discussed.

Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection.

Globally, the emergence of the coronavirus disease (COVID-19) has had a significant impact on life. The need for ongoing SARS-CoV-2 screening employing inexpensive and quick diagnostic approaches is undeniable, given the ongoing pandemic and variations in vaccine administration in resource-constrained regions. This study presents results as proof of concept to use hybridization chain reaction (HCR) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a complex for detecting SARS-CoV-2. HCR hairpin probes were designed using the NUPACK web-based program and further used to amplify the SARS-CoV-2 N gene in archived nasopharyngeal samples. The results were visualized using agarose gels and CRISPR Cas12a-based lateral flow strips. The assay was evaluated using the gold standard, real-time polymerase chain reaction (RT-PCR), as recommended by the World Health Organization (WHO). The results show the comparative efficiency of HCR to RT-PCR. This study shows that HCR and CRISPR are viable alternatives for diagnosing SARS-CoV-2 in samples.

Evaluation of Simplified HCV Diagnostics in HIV/HCV Co-Infected Patients in Myanmar.

To evaluate a decentralised testing model and simplified treatment protocol of hepatitis C virus (HCV) infection to facilitate treatment scale-up in Myanmar, this prospective, observational study recruited HIV-HCV co-infected outpatients receiving sofosbuvir/daclatasvir in Yangon, Myanmar. The study examined the outcomes and factors associated with a sustained virological response (SVR). A decentralised "hub-and-spoke" testing model was evaluated where fingerstick capillary specimens were transported by taxi and processed centrally. The performance of the Xpert HCV VL Fingerstick Assay in detecting HCV RNA was compared to the local standard of care ( plasma HCV RNA collected by venepuncture). Between January 2019 and February 2020, 162 HCV RNA-positive individuals were identified; 154/162 (95%) initiated treatment, and 128/154 (84%) returned for their SVR12 visit. A SVR was achieved in 119/154 (77%) participants in the intent-to-treat population and 119/128 (93%) participants in the modified-intent-to-treat population. Individuals receiving an antiretroviral therapy were more likely to achieve a SVR (with an odds ratio (OR) of 7.16, 95% CI 1.03-49.50), while those with cirrhosis were less likely (OR: 0.26, 95% CI 0.07-0.88). The sensitivity of the Xpert HCV VL Fingerstick Assay was 99.4% (95% CI 96.7-100.0), and the specificity was 99.2% (95% CI 95.9-99.9). A simplified treatment protocol using a hub-and-spoke testing model of fingerstick capillary specimens can achieve an SVR rate in LMIC comparable to well-resourced high-income settings.

Evaluation of Urine L-FABP Point of Care Kit in the Philippines as Predictive Marker of Clinical Severity of COVID-19 (EPOCH COVID study)

Background: The search for simple clinical and laboratory markers to help predict the clinical severity of patients presenting with COVID-19 has prompted this study to look at the predictive value of urine L-FABP (Liver Type-Fatty Acid Binding Protein) point-of-care test kit at the initial presentation of COVID-19 patients to the hospital. Method(s): The validation study prospectively included 109 consecutive patients with mild to moderate COVID-19, mean age of 52.2 years (range 19-84) presenting at the Emergency Rooms of 4 participating Metro-Manila hospitals from February to April 2021, with available data for analysis for 103 patients. Urine L-FABP POC (Point-of-Care) test and other clinical parameters and the level of severity of COVID-19 were determined at Day 0, Day 4 and Day 7. Computations for Sensitivity, Specificity, Positive and Negative Predictive values and Likelihood ratios were performed Results: Twenty-three patients tested positive for urine L-FABP, out of the 103 patients analyzed, while 80 tested negative. Of the 23 patients who tested positive for urine L-FABP, 6 has progressed in severity, while 17 did not progressed. Of the 80 patients who tested negative for urine L-FABP, 13 progressed, while 67 did not progressed in severity. Giving a Sensitivity of 31.58%, Specificity of 79.76%, Positive predictive value of 26.09%, Negative predictive value of 83.75%. Combining urine L-FABP and initial clinical parameters like SIRS (Systemic Inflammatory Response Syndrome) criteria to predict progression of severity yielded a higher Specificity of 91.67 % and Negative Predictive value of 84.62%. Conclusion(s): The study shows the utility of initial urine L-FABP POC test as a negative screening test in triaging adult patients presenting to the ER with mild to moderate COVID-19. Patients at the ER with a negative urine L-FABP test, will most likely not progressed to severe COVID-19. Combining clinical parameters like SIRS Criteria with the urine L-FABP result can increase the negative predictive value.Copyright © The PHILIPPINE JOURNAL OF INTERNAL MEDICINE is a peer reviewed journal and a copyrighted publication of the Philippine College of Physicians.

Health Care Monitoring Application using IoT Devices

Nowadays, health monitoring is crucial, especially monitoring the temperature and heartbeat of the patient in Covid / non-Covid situations. Continued monitoring of the patient is not a possible and tedious job. IoT plays a critical role in Hospitals where patients are in Intensive Care Units (ICU), and patients are treated at home (isolation points). The devices receive data continuously and monitor by the doctors remotely. This paper presents temperature and heartbeat monitoring using Internet Of Things (IoT) devices with an algorithm to capture data from devices and sends it to computer devices at a reasonable cost. Proof Of Concept has been created with the help of an Arduino board, Pulse Sensor, Temperature Sensor, Breadboard, ESP8266 Wi-Fi module, and Liquid Crystal Display (LCD). The IoT devices capture data from different devices (patient health data) in real time. The Health Care Monitoring (HCM) Application builds using microservices architecture, runs on top of the Thingspeak data, and sends notifications to the doctors if there is an emergency. The doctors can act according to rather than monitor continuously. This model eliminates manual intervention for taking the reading from time to time. © 2022 IEEE.

Evaluation of the Visby medical COVID-19 point of care nucleic acid amplification test.

Rapid and widespread diagnostic testing is critical to providing timely patient care and reducing transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Recently, the Visby Medical COVID-19 point of care (POC) test was granted emergency use authorization (EUA) for qualitative detection of SARS-CoV-2 nucleic acid at the point of care. We evaluated its performance characteristics using residual specimens (n = 100) collected from Mayo Clinic patients using nasopharyngeal (NP) swabs and placed in viral transport media (VTM). The same specimen was tested using both the laboratory reference method (RT-qPCR) and Visby test. The reference methods utilized included a laboratory developed test with EUA (Mayo Clinic Laboratories, Rochester, MN) using the TaqMan assay on a Roche Light Cycler 480 or a commercially available EUA platform (cobas® SARS-CoV-2; Roche Diagnostics, Indianapolis, IN). Positive, negative, and overall percent agreement between the Visby COVID-19 test and the reference method were calculated. Additionally, the limit of detection (LoD) claimed by the manufacturer (1112 copies/mL) was verified with serial dilutions of heat inactivated virus. The Visby COVID-19 test correctly identified 29/30 positive samples and 69/70 negative samples, resulting in an overall concordance of 98.0%, positive percent agreement of 96.7%, and negative percent agreement of 98.6%. The abbreviated LoD experiment showed that the analytical sensitivity of the method is as low as or lower than 500 copies/mL. Our study demonstrated that Visby COVID-19 is well-suited to address rapid SARS-CoV-2 testing needs. It has high concordance with central laboratory-based RT-qPCR methods, a low rate of invalid results, and superior analytical sensitivity to some other EUA POC devices.

Evaluation of Abbott ID NOW COVID-19 POC test performance characteristics and integration in the regional health network workflows to improve health care delivery.

With the recent global surge of SARS-CoV-2 Delta variant, there continues to be high demand for COVID-19 diagnostic testing. Abbott ID NOW is a rapid, CLIA-waived, COVID-19 diagnostic test ideally suited for use in urgent care settings or where access to diagnostic testing is limited. In this study we describe the results of rigorous validation of ID NOW and post-implementation study of POC test utilization patterns within community hospitals and clinics. Performance of ID NOW was validated by comparison of the results from 207 consecutive, paired, specimens tested on the ID NOW and on the m2000/Alinity m platforms. Once validated, ID NOW devices were placed for clinical use at four regional hospitals and clinics. We found that the ID NOW and m2000/Alinity m positive and negative percent agreement were 94.5% (95% CI, 85.1% to 98.1%) and 99.3% (95% CI, 96.4% to 99.9%), respectively. As of August 2021, a total of 2,301 tests were performed by ID NOW at individual regional network sites. The population tested consisted of 55.5% White and 42.9% Black patients, with Black patients presenting predominantly in the hospitals, while White patients were more evenly distributed between hospital and clinic sites. Disease prevalence observed among patients tested by ID NOW (12.3%) was aligned with overall prevalence seen at regional sites (11.3%). In summary, the ID NOW test can provide rapid and accurate results in a variety of near-to-patient and POC settings. If used correctly, it could serve as a valuable diagnostic tool to enable equal access to care and improve healthcare delivery within large health network systems.

A Comparative Study of Voltammetric vs Impedimetric Immunosensor for Rapid SARS-CoV-2 Detection at the Point-of-care.

Here, a novel biosensing platform for the detection of SARS-CoV-2 usable both at voltammetric and impedimetric mode is reported. The platform was constructed on a multi-walled carbon nanotubes (MWCNTs) screen-printed electrode (SPE) functionalized by methylene blue (MB), antibodies against SARS-CoV-2 spike protein (SP), a bioactive layer of chitosan (CS) and protein A (PrA). The voltammetric sensor showed superior performances both in phosphate buffer solution (PBS) and spiked-saliva samples, with LOD values of 5.0±0.1 and 30±2.1 ng/mL, compared to 20±1.8 and 50±2.5 ng/mL for the impedimetric sensor. Moreover, the voltammetric immunosensor was tested in real saliva, showing promising results.

Point-of-Care Biosensors for Healthcare Applications

In the healthcare sector, biosensors have been extensively used to detect pathogens, antigens, and biomarkers for different diseases/ailments from various biological samples like blood serum, plasma, urine, saliva, faecal matter, etc. Point-of-care (POC) biosensors are scaled down to compact devices that can detect diseases next to the patient to reduce the therapeutic turnaround time. Determination of blood sugar level for diabetes monitoring is the most widely used and commercially available, self-usable POC biosensor. Although biosensors exist for various diseases, there is still a challenge of making them POC because of characteristic requirements of the bioreceptors, such as the storage conditions, and fabrication techniques. Cardiovascular diseases, neural disease (stress), kidney disease, urinary tract infection, and other viral infections are some diseases that can be detected using a biosensor. POC biosensors are available for detecting some of these diseases. This book chapter discusses different POC biosensors for all the prominent diseases and conditions. A good number of rapid POC devices for mass testing and detection of coronavirus disease 2019 (COVID-19) were quickly developed, which helped in controlling the pandemic. Handheld electronic device systems to display the outputs and results of the biosensors are also available for many of the biosensors. The advancement of the Internet of things (IoT) made these biosensor devices linkable with a smartphone to make delivery of results possible for the doctors to analyse. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2022.

Detection and Diagnosis Technologies for SARS-CoV-2

Detection and diagnosis platforms play key roles in early warning, outbreak control and exit strategy for any pandemic, and they are especially pertinent for the Coronavirus disease 2019 (COVID-19) pandemic. The challenges posed by the speed and extent of severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) spread around the globe also offered unprecedented opportunities for the development and deployment of novel strategies and products - not only vaccines and therapeutics, but also diagnostics. This chapter provides a brief summary of the vast array of molecular, serological, cell-based and other diagnostic tools for the specific detection of SARS-CoV-2 infections and immune responses. The focus is on the principles and applications of each platform, while detailed protocols can be found in the cited references. © 2023 by World Scientific Publishing Co. Pte. Ltd.

Perspective on Aggregation-induced Emission (AIE) Materials for Pathogen Detection

The rapid and accurate identification of pathogens plays a crucial role in clinical practice, which helps to prevent, control, and treat pathogenic infections at the initial stage. The current available technologies for pathogen detection appear to be inadequate in dealing with cases such as COVID-19. More importantly, the frequent emergence of drug-resistant bacteria is gradually rendering the existing therapeutic options ineffective. Efforts are urgently required to focus on the development of diagnostic systems for point-of-care (POC) detection and high-throughput pathogen identification. Since 2001, a new class of aggregation-induced emission luminogens (AIEgens) with good photostability, high sensitiv-ity, and improved signal-to-noise ratio has emerged as powerful fluorescent tools for various biosensing and cell imaging. Based on the unique fluorescence of AIEgens that becomes stronger upon aggregation, naked-eye detection in turn-on mode has gained a speedy development. A timely overview can not only provide a summary of the advances and challenges of AIEgens in pathogen detection but also offer sys-tematic ideas for future developments. There are also expectations for in-depth interdisciplinary research in the field of analytical chemistry and microbiology. © 2023 Bentham Science Publishers.

Analysis of Quality Indicators of the Pre-Analytical Phase on Blood Gas Analyzers, Point-Of-Care Analyzer in the Period of the COVID-19 Pandemic.

AIM OF THE STUDY: We evaluated and compared blood gas analysis (EGA) non-conformities (NC) considered operator-dependent performed in Point-Of-Care (POC) analyzer as quality indicators (IQ) of the pre-analytical phase. To this end, four different NC registered in the resuscitation departments of the Hospital Polyclinic Bari from the beginning of the pandemic (March 2020) until February 2022 were evaluated. The results obtained were compared with those recorded in the pre-COVID period (March 2018-February 2020) to check if there were differences in number and type. MATERIAL AND METHODS: GEM 4000 series blood gas analyzers (Instrumentation Laboratory, Bedford, MA, United States) are installed with integrated Intelligent Quality Management (iQM®), which automatically identify and log pre-analytical errors. All blood gas analyzers are connected to the company intranet and interfaced with the GEM Web Plus (Werfen Instrumentation Laboratory, Bedford, MA, United States) data management information system, which allows the core laboratory to remotely supervise all decentralized POC stations. The operator-dependent process NC were expressed in terms of absolute and relative proportions (percentiles and percentage changes). For performance evaluation, the Mann-Whitney U test, Chi-squared test and Six-Sigma Metric calculation for performance classification were performed. RESULTS: In the COVID period, 31,364 blood gas tests were performed vs. 16,632 tests in the pre-COVID period. The NC related to the suitability of the EGA sample and manageable by the operators were totals of 652 (3.9%) and 749 (2.4%), respectively, in the pre-COVID and COVID periods. The pre-analytical phase IQs used did not show statistically significant differences in the two periods evaluated. The Sigma evaluation did not show an increase in error rates. CONCLUSIONS: Considering the increase in the number of EGAs performed in the two periods, the training procedures performed by the core laboratory staff were effective; the clinical users of the POC complied with the indications and procedures shared with the core laboratory without increasing the operator-dependent NCs. Furthermore, the core laboratory developed monitoring activities capable of guaranteeing the maintenance of the pre-analytical quality.

A fully-enclosed prototype 'pen' for rapid detection of SARS-CoV-2 based on RT-RPA with dipstick assay at point-of-care testing.

A fully-enclosed prototype 'pen' for rapid detection of SARS-CoV-2 based on reverse transcriptase isothermal recombinase polymerase amplification (RT-RPA) with dipstick assay was developed. The integrated handheld device, consisting of amplification, detection and sealing modules, was developed to perform rapid nucleic acid amplification and detection under a fully enclosed condition. After RT-RPA amplification with a metal bath or a normal PCR instrument, the amplicons were mixed with dilution buffer prior to being detected on a lateral flow strip. To avoid aerosol contamination causing false-positive, from amplification to final detection, the detection 'pen' had been enclosed to isolate from the environment. With colloidal gold strip-based detection, the detection results could be directly observed by eyes. By cooperating with other inexpensive and rapid methods for POC nucleic acid extraction, the developed 'pen' could detect COVID-19 or other infectious diseases in a convenient, simple and reliable way.

Validation of Rapid and Economic Colorimetric Nanoparticle Assay for SARS-CoV-2 RNA Detection in Saliva and Nasopharyngeal Swabs.

Even with the widespread uptake of vaccines, the SARS-CoV-2-induced COVID-19 pandemic continues to overwhelm many healthcare systems worldwide. Consequently, massive scale molecular diagnostic testing remains a key strategy to control the ongoing pandemic, and the need for instrument-free, economic and easy-to-use molecular diagnostic alternatives to PCR remains a goal of many healthcare providers, including WHO. We developed a test (Repvit) based on gold nanoparticles that can detect SARS-CoV-2 RNA directly from nasopharyngeal swab or saliva samples with a limit of detection (LOD) of 2.1 × 105 copies mL-1 by the naked eye (or 8 × 104 copies mL-1 by spectrophotometer) in less than 20 min, without the need for any instrumentation, and with a manufacturing price of <$1. We tested this technology on 1143 clinical samples from RNA extracted from nasopharyngeal swabs (n = 188), directly from saliva samples (n = 635; assayed by spectrophotometer) and nasopharyngeal swabs (n = 320) from multiple centers and obtained sensitivity values of 92.86%, 93.75% and 94.57% and specificities of 93.22%, 97.96% and 94.76%, respectively. To our knowledge, this is the first description of a colloidal nanoparticle assay that allows for rapid nucleic acid detection at clinically relevant sensitivity without the need for external instrumentation that could be used in resource-limited settings or for self-testing.

Multiplexed Ultrasensitive Sample-to-Answer RT-LAMP Chip for the Identification of SARS-CoV-2 and Influenza Viruses.

Prompt on-site diagnosis of SARS-CoV-2 with other respiratory infections would have minimized the global impact of the COVID-19 pandemic through rapid, effective management. However, no such multiplex point-of-care (POC) chip has satisfied a suitable sensitivity of gold-standard nucleic acid amplification tests (NAATs). Here, we present a rapid multiplexed ultrasensitive sample-to-answer LAMP (MUSAL) chip operated by simple LED-driven photothermal amplification to detect six targets from single-swab sampling. First, our MUSAL chip allows ultrafast on-chip sample preparation with ∼500-fold preconcentration at a rate of 1.2 mL min-1 . Second, our chip enables contamination-free amplification using autonomous target elution into on-chip reagents by photothermal activation. Finally, our chip accomplishes multiplexed on-chip diagnostics of SARS-CoV-2 and influenza viruses with a limit of detection (LoD) of 0.5 copies µL-1 . Our rapid, ultra-sensitive, cost-effective sample-to-answer chip with a multiplex capability will allow timely management of various pandemics situations we may face shortly. This article is protected by copyright. All rights reserved.

ICT Enabled Disease Diagnosis, Treatment and Management-A Holistic Cost-Effective Approach Through Data Management and Analysis in UAE and India.

This concept paper addresses specific challenges identified in the UN 2030 Agenda Sustainable Development Goals (SDG) as well as the National Health Policy of India (NHP-India) and the Ministry of Health Policy of UAE (MHP-UAE). This policy calls for a digital health technology ecosystem. SDG Goal 1 and its related objectives are conceptualized which serves as the foundation for Virtual Consultations, Tele-pharmacy, Virtual Storage, and Virtual Community (VCom). SDG Goals 2 and 3 are conceptualized as Data Management & Analytical (DMA) Architecture. Individual researchers and health care professionals in India and the UAE can use DMA to uncover and harness PHC and POC data into practical insights. In addition, the DMA would provide a set of core tools for cross-network initiatives, allowing researchers and other users to compare their data with DMA data. In rural, urban, and remote populations of the UAE and India, the concept augments the PHC system with ICT-based interventions. The ICT-based interventions may improve patient health outcomes. The open and flexible design allows users to access various digital materials. Extendable data/metadata format, scalable architecture for petabyte-scale federated discovery. The modular DMA is designed using existing technology and resources. Public health functions include population health assessment, policy development, and monitoring policy implementation. PHC and POC periodically conduct syndromic surveillance to identify population risk patterns. In addition, the PHC and POC deploy medical and non-medical preventive measures to prevent disease outbreaks. To assess the impact of social and economic factors on health, epidemiologists must first understand diseases. Improved health due to compliance with holistic disease treatment plans and access to scientific health information.

CRISPR-Based Diagnostics: Challenges and Potential Solutions toward Point-of-Care Applications.

The COVID-19 pandemic has challenged the conventional diagnostic field and revealed the need for decentralized Point of Care (POC) solutions. Although nucleic acid testing is considered to be the most sensitive and specific disease detection method, conventional testing platforms are expensive, confined to central laboratories, and are not deployable in low-resource settings. CRISPR-based diagnostics have emerged as promising tools capable of revolutionizing the field of molecular diagnostics. These platforms are inexpensive, simple, and do not require the use of special instrumentation, suggesting they could democratize access to disease diagnostics. However, there are several obstacles to the use of the current platforms for POC applications, including difficulties in sample processing and stability. In this review, we discuss key advancements in the field, with an emphasis on the challenges of sample processing, stability, multiplexing, amplification-free detection, signal interpretation, and process automation. We also discuss potential solutions for revolutionizing CRISPR-based diagnostics toward sample-to-answer diagnostic solutions for POC and home use.

Prospective evaluation of thoracic diseases using a compact flat-panel detector spiral computed tomographic scanner.

Objective: To prospectively evaluate the image quality and diagnostic performance of a compact flat-panel detector (FD) scanner for thoracic diseases compared to a clinical CT scanner. Materials and methods: The institutional review board approved this single-center prospective study, and all participants provided informed consent. From December 2020 to May 2021, 30 patients (mean age, 67.1 ± 8.3 years) underwent two same-day low-dose chest CT scans using clinical state-of-art and compact FDCT scanners. Image quality was assessed visually and quantitatively. Two readers evaluated the diagnostic performance for nodules, parenchymal opacifications, bronchiectasis, linear opacities, and pleural abnormalities in 40 paired CT scans. The other 20 paired CT scans were used to examine the agreement of semi-quantitative CT scoring regarding bronchiectasis, bronchiolitis, nodules, airspace consolidations, and cavities. Results: FDCT images had significantly lower visual image quality than clinical CT images (all p < 0.001). The two CT image sets showed no significant differences in signal-to-noise and contrast-to-noise ratios (56.8 ± 12.5 vs. 57.3 ± 15.2; p = 0.985 and 62.9 ± 11.7 vs. 60.7 ± 16.9; p = 0.615). The pooled sensitivity was comparable for nodules, parenchymal opacifications, linear opacities, and pleural abnormalities (p = 0.065-0.625), whereas the sensitivity was significantly lower in FDCT images than in clinical CT images for micronodules (p = 0.007) and bronchiectasis (p = 0.004). The specificity was mostly 1.0. Semi-quantitative CT scores were similar between the CT image sets (p > 0.05), and intraclass correlation coefficients were around 0.950 or higher, except for bronchiectasis (0.869). Conclusion: Compact FDCT images provided lower image quality but comparable diagnostic performance to clinical CT images for nodules, parenchymal opacifications, linear opacities, and pleural abnormalities.

CRISPR-Cas12a coupled with universal gold nanoparticle strand-displacement probe for rapid and sensitive visual SARS-CoV-2 detection.

Point of care (POC) diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are particularly significant for preventing transmission of coronavirus disease 2019 (COVID-19) by any user at any given time and place. CRISPR/Cas-assisted SARS-CoV-2 assays are viewed as supplemental to RT-PCR due to simple operation, convenient use and low cost. However, most current CRISPR molecular diagnostics based on fluorescence measurement increased the difficulty of POC test with need of the additional light sources. Some instrument-free visual detection with the naked eye has limitations in probe universality. Herein, we developed a universal, rapid, sensitive and specific SARS-CoV-2 POC test that combines the outstanding DNase activity of Cas12a with universal AuNPs strand-displacement probe. The oligo trigger, which is the switch the AuNPs of the strand-displacement probe, is declined as a result of Cas12a recognition and digestion. The amount of released AuNPs produced color change which can be visual with the naked eye and assessed by UV-Vis spectrometer for quantitative detection. Furthermore, a low-cost hand warmer is used as an incubator for the visual assay, enabling an instrument-free, visual SARS-CoV-2 detection within 20 min. A real coronavirus GX/P2V instead of SARS-CoV-2 were chosen for practical application validation. After rapid virus RNA extraction and RT-PCR amplification, a minimum of 2.7 × 102 copies/mL was obtained successfully. The modular design can be applied to many nucleic acid detection applications, such as viruses, bacteria, species, etc., by simply modifying the crRNA, showing great potential in POC diagnosis.

Using human factors principles to redesign a 3D lab workflow during the COVID-19 pandemic.

BACKGROUND: Like most hospitals, our hospital experienced COVID-19 pandemic-related supply chain shortages. Our additive manufacturing lab's capacity to offset these shortages was soon overwhelmed, leading to a need to improve the efficiency of our existing workflow. We undertook a work system analysis guided by the Systems Engineering Initiative for Patient Safety (SEIPS) construct which is based on human factors and quality improvement principles. Our objective was to understand the inefficiencies in project submission, review, and acceptance decisions, and make systematic improvements to optimize lab operations. METHODS: Contextual inquiry (interviews and workflow analysis) revealed suboptimal characteristics of the system, specifically, reliance on a single person to facilitate work and, at times, fractured communication with project sponsors, with root causes related to the project intake and evaluation process as identified through SEIPS tools. As interventions, the analysis led us to: 1) enhance an existing but underused project submission form, 2) design and implement an internal project scorecard to standardize evaluation of requests, and 3) distribute the responsibility of submission evaluation across lab members. We implemented these interventions in May 2021 for new projects and compare them to our baseline February 1, 2018 through - April 30, 2021 performance (1184 days). RESULTS: All project requests were submitted using the enhanced project submission form and all received a standardized evaluation with the project scorecard. Prior to interventions, we completed 35/79 (44%) of projects, compared to 12/20 (60%) of projects after interventions were implemented. Time to review new submissions was reduced from an average of 58 days to 4 days. A more distributed team responsibility structure permitted improved workflow with no increase in staffing, allowing the Lab Manager to devote more time to engineering rather than administrative/decision tasks. CONCLUSIONS: By optimizing our workflows utilizing a human factors approach, we improved the work system of our additive manufacturing lab to be responsive to the urgent needs of the pandemic. The current workflow provides insights for labs aiming to meet the growing demand for point-of-care manufacturing.