Klebsiella pneumoniae poses a significant threat to global public health. Traditional clinical diagnostic methods, such as bacterial culture and microscopic identification, are not suitable for point-of-care testing. In response, based on the suboptimal protospacer adjacent motifs, this study develops an extraction-free one-pot assay, named EXORCA (EXtraction-free One-pot RPA-CRISPR/Cas12a assay), designed for the immediate, sensitive and efficient detection of K. pneumoniae. The EXORCA assay can be completed within approximately 30 min at a constant temperature and allows for the visualization of results either through a fluorescence reader or directly by the naked eye under blue light. The feasibility of the assay was evaluated using twenty unextracted clinical samples, achieving a 100% (5/5) positive predictive value and a 100% (15/15) negative predictive value in comparison to qPCR. These results suggest that the EXORCA assay holds significant potential as a point-of-care testing tool for the rapid identification of pathogens, such as K. pneumonia.

BACKGROUND: Temperature sensing is commonly used in point-of-care (POC) detection technologies, yet the portability and convenience of use are frequently compromised by the complexity of thermosensitive processes and signal transduction. Especially, multi-step target recognition reactions and temperature measurement in the reaction vessel present challenges in terms of stability and integration of detection devices. To further combine photothermal reaction and signal readout in one assay, these two processes enable to be integrated into miniaturized microfluidic chips, thereby facilitating photothermal sensing and achieving a simple visual temperature sensing as POC detection.
RESULTS: A copper ion (Cu2+)-catalyzed photothermal sensing system integrated onto a microfluidic distance-based analytical device (μDAD), enabling the visual, portable, and sensitive quantitative detection of multiple targets, including ascorbic acid, glutathione, and alkaline phosphatase (ALP). The polydopamine nanoparticles (PDA NPs) were synthesized by the regulation of free Cu2+ through redox or coordination reactions, facilitating the transduction of distinct photothermal response signals and providing the versatile Cu2+-responsive sensing systems. Promoted by integration with a photothermal μDAD, the system combines PDA\'s photothermal responsiveness and thermosensitive gas production of ammonium bicarbonate for improved sensitivity of ALP detection, reaching the detection limit of 9.1 mU/L. The system has successfully achieved on-chip detection of ALP with superior anti-interference capability and recoveries ranging from 96.8 % to 104.7 %, alongside relative standard deviations below 8.0 %.
UNASSIGNED: The μDAD design accommodated both the photothermal reaction of PDA NPs and thermosensitive gas production reaction, achieving the rapid sensing of visual distance signals. The μDAD-based Cu2+-catalyzed photothermal sensing system holds substantial potential for applications in biochemical analysis and clinical diagnostics, underscored by the versatile Cu2+ regulation mechanism for a broad spectrum of biomarkers.

BACKGROUND: A novel single-use, analyser-free, molecular point-of-care test for SARS-CoV-2 (Veros COVID-19 test, Sherlock Biosciences) could reduce time to results and improve patient care and flow in the emergency department (ED) but its performance in this setting is unknown.
METHODS: Adults aged ≥18 years presenting to Southampton General Hospital (UK) with suspected COVID-19 were tested with the Veros COVID-19 test in addition to standard of care near-patient PCR. Measures of diagnostic accuracy were calculated for the Veros COVID-19 test stratified by Ct value. Discrepant results underwent viral culture.
RESULTS: Between Jan 16 and May 2 2023, 400 patients were enrolled with a median (IQR) age of 60 (34-77) and 141 (35·3%) were SARS-CoV-2 positive by PCR. The Veros test gave valid results on the first test in 384 (96·0%) and sensitivity and specificity were 127/141 (90·1%, 95%CI 83·9-94·5) and 258/259 (99·6%, 95%CI 97·9-100) overall. For those with high or moderate viral load (Ct ≤30), sensitivity was 125/129 (96·9%, 95%CI 92·3-99·2). One (7·1%) of 14 PCR positive/Veros test negative samples was culture positive. Median (IQR) time from sample collection to result was 19 (18-20) mins with the Veros test versus 73 (59-92) mins with PCR (p<0·0001).
CONCLUSIONS: The Veros COVID-19 test generated results in near real-time, around 1hour sooner than rapid, near-patient, analyser-based PCR and accuracy was excellent for samples with moderate and high viral loads. The Veros test represents a step-change in molecular diagnostics for infection and could significantly reduce time to results and improve patient management in EDs and other settings.
BACKGROUND: Sherlock Biosciences DATA SHARING: All de-identified participant data analysed and presented in this study are available from the corresponding author following publication, on reasonable request.

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Background. The COVID-19 pandemic demonstrated a need for robust SARS-CoV-2 test evaluation infrastructure to underpin biosecurity and protect the population during a pandemic health emergency.Gap statement. The first generation of rapid antigen tests was less accurate than molecular methods due to their inherent sensitivity and specificity shortfalls, compounded by the consequences of self-testing. This created a need for more accurate point-of-care SARS-CoV-2 detection methods.Aim. Here we present the lessons-learned during the COVID-19 emergency response in Western Australia including the detailed set-up, evaluation and operation of rapid antigen test in a state-run drive-through sample collection service during the COVID-19 pandemic after the strict border shutdown ended.Methods. We report a conformity assessment of a novel, second-generation rapid antigen test (Virulizer) comprising a technician-operated rapid lateral flow immunoassay with fluorescence-based detection.Results. The Virulizer rapid antigen test demonstrated up to 100% sensitivity (95% CI: 61.0-100%), 91.94% specificity (95% CI: 82.5-96.5%) and 92.65% accuracy when compared to a commercial PCR assay method. Wide confidence intervals in our series reflect the limits of small sample size. Nevertheless, the Virulizer assay performance was well-suited to point-of-care screening for SARS-CoV-2 in a drive-through clinic setting.Conclusion. The adaptive evaluation process necessary under changing pandemic conditions enabled assessment of a simple sample collection and point-of-care testing process, and showed how this system could be rapidly deployed for SARS-CoV-2 testing, including to regional and remote settings.

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In this dual-center study, we assessed the BioHermes A1C EXP M13 system for point-of-care (POC) HbA1c testing against two NGSP-certified HPLC instruments, the Bio-Rad D100 and Tosoh G8. Analyzing 605 samples, we evaluated the A1C EXP\'s reproducibility, sensitivity, specificity and impact of anemia on HbA1c measurements. The device showed excellent reproducibility with CVs under 2.4% and high sensitivity and specificity for diabetes diagnosis-98.1% and 96.8% against D100, and 97.1% and 96.7% against G8. Passing-Bablok regression confirmed a close correlation between A1C EXP and the HPLC instruments, with equations y = 0.10625 + 0.9688x (D100) and y = 0.0000 + 0.1000x (G8), and Bland-Altman plots indicated mean relative differences of -1.4% (D100) and -0.4% (G8). However, in anemic samples, A1C EXP showed a negative bias compared to HPLC devices, suggesting that anemia may affect the accuracy of HbA1c results. The study indicates that A1C EXP is a reliable POC alternative to laboratory assays, albeit with considerations for anemic patients.

Infectious diseases significantly impact global health, necessitating prompt diagnosis to mitigate life-threatening sepsis risk. Identifying patients at risk of severe neurological complications from enterovirus infections is challenging due to nonspecific initial presentations. Point-of-care testing (POCT) has emerged as a transformative tool, with low-cost lateral-flow colorimetric assays showing promise in deployable POCT devices. We developed a PCT/IL-6 rapid diagnostic system integrating lateral flow assay (LFA) test strips and a portable optical spectrum reader, allowing simultaneous semi-quantitative measurement of serum PCT and IL-6 within 30 min at the point of care. The system demonstrated a strong correlation with traditional ELISA and effectively differentiated severe pediatric enterovirus cases using serum samples. IL-6 showed superior discriminatory ability over PCT in identifying patients with severe neurological complications. This novel diagnostic platform holds great potential for early sepsis recognition and infectious disease management, especially in resource-limited settings.

Molecularly imprinted membranes (MIMs) have been a focal research interest since 1990, representing a breakthrough in the integration of target molecules into membrane structures for cutting-edge sensing applications. This paper traces the developmental history of MIMs, elucidating the diverse methodologies employed in their preparation and characterization on two-dimensional solid-supported substrates. We then explore the principles and diverse applications of MIMs, particularly in the context of emerging technologies encompassing electrochemistry, surface-enhanced Raman scattering (SERS), surface plasmon resonance (SPR), and the quartz crystal microbalance (QCM). Furthermore, we shed light on the unique features of ion-sensitive field-effect transistor (ISFET) biosensors that rely on MIMs, with the notable advancements and challenges of point-of-care biochemical sensors highlighted. By providing a comprehensive overview of the latest innovations and future trajectories, this paper aims to inspire further exploration and progress in the field of MIM-driven sensing technologies.

A novel, portable deep learning-assisted smartphone-based electrochemiluminescence (ECL) cost-effective (~10$) sensing platform was developed and used for selective detection of lactate. Low-cost, fast prototyping screen printing and wax printing methods with paper-based substrate were used to fabricate miniaturized single-pair electrode ECL platforms. The lab-made 3D-printed portable black box served as a reaction chamber. This portable platform was integrated with a smartphone and a buck-boost converter, eliminating the need for expensive CCD cameras, photomultiplier tubes, and bulky power supplies. This advancement makes this platform ideal for point-of-care testing applications. Foremost, the integration of a deep learning approach served to enhance not just the accuracy of the ECL sensors, but also to expedite the diagnostic procedure. The deep learning models were trained (3600 ECL images) and tested (900 ECL images) using ECL images obtained from experimentation. Herein, for user convenience, an Android application with a graphical user interface was developed. This app performs several tasks, which include capturing real-time images, cropping them, and predicting the concentration of required bioanalytes through deep learning. The device\'s capability to work in a real environment was tested by performing lactate sensing. The fabricated ECL device shows a good liner range (from 50 µM to 2000 µM) with an acceptable limit of detection value of 5.14 µM. Finally, various rigorous analyses, including stability, reproducibility, and unknown sample analysis, were conducted to check device durability and stability. Therefore, the developed platform becomes versatile and applicable across various domains by harnessing deep learning as a cutting-edge technology and integrating it with a smartphone.