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1. New technologies and reagents in lateral flow assay (LFA) designs for enhancing accuracy and sensitivity

   https://doi.org/10.1039/D3AY00844D  

   Dey, Mohan K; Iftesum, Maria; Devireddy, Ram; Gartia, Manas R (September 2023, Analytical Methods)

   Lateral flow assays (LFAs) are a popular method for quick and affordable diagnostic testing because they are easy to use, portable, and user-friendly. However, LFA design has always faced challenges regarding sensitivity, accuracy, and complexity of the operation. By integrating new technologies and reagents, the sensitivity and accuracy of LFAs can be improved while minimizing the complexity and potential for false positives. Surface enhanced Raman spectroscopy (SERS), photoacoustic techniques, fluorescence resonance energy transfer (FRET), and the integration of smartphones and thermal readers can improve LFA accuracy and sensitivity. To ensure reliable and accurate results, careful assay design and validation, appropriate controls, and optimization of assay conditions are necessary. Continued innovation in LFA technology is crucial to improving the reliability and accuracy of rapid diagnostic testing and expanding its applications to various areas, such as food testing, water quality monitoring, and environmental testing. 

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2. Monocular depth estimation based on deep learning for intraoperative guidance using surface-enhanced Raman scattering imaging

   https://doi.org/10.1364/PRJ.536871  

   Juhong, Aniwat; Li, Bo; Liu, Yifan; Yao, Cheng-You; Yang, Chia-Wei; Atique_Ullah, A_K M; Liu, Kunli; Lewandowski, Ryan P; Harkema, Jack R; Agnew, Dalen W; et al (January 2025, Photonics Research)

   Imaging of surface-enhanced Raman scattering (SERS) nanoparticles (NPs) has been intensively studied for cancer detection due to its high sensitivity, unconstrained low signal-to-noise ratios, and multiplexing detection capability. Furthermore, conjugating SERS NPs with various biomarkers is straightforward, resulting in numerous successful studies on cancer detection and diagnosis. However, Raman spectroscopy only provides spectral data from an imaging area without co-registered anatomic context. This is not practical and suitable for clinical applications. Here, we propose a custom-made Raman spectrometer with computer-vision-based positional tracking and monocular depth estimation using deep learning (DL) for the visualization of 2D and 3D SERS NPs imaging, respectively. In addition, the SERS NPs used in this study (hyaluronic acid-conjugated SERS NPs) showed clear tumor targeting capabilities (target CD44 typically overexpressed in tumors) by anex vivoexperiment and immunohistochemistry. The combination of Raman spectroscopy, image processing, and SERS molecular imaging, therefore, offers a robust and feasible potential for clinical applications. 

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3. Smartphone-read phage lateral flow assay for point-of-care detection of infection

   https://doi.org/10.1039/d2an01499h  

   Chabi, Maede; Vu, Binh; Brosamer, Kristen; Smith, Maxwell; Chavan, Dimple; Conrad, Jacinta C.; Willson, Richard C.; Kourentzi, Katerina (February 2023, The Analyst)

   The COVID-19 pandemic has highlighted the urgent need for sensitive, affordable, and widely accessible testing at the point of care. Here we demonstrate a new, universal LFA platform technology using M13 phage conjugated with antibodies and HRP enzymes that offers high analytical sensitivity and excellent performance in a complex clinical matrix. We also report its complete integration into a sensitive chemiluminescence-based smartphone-readable lateral flow assay for the detection of SARS-CoV-2 nucleoprotein. We screened 84 anti-nucleoprotein monoclonal antibody pairs in phage LFA and identified an antibody pair that gave an LoD of 25 pg mL −1 nucleoprotein in nasal swab extract using a FluorChem gel documentation system and 100 pg mL −1 when the test was imaged and analyzed by an in-house-developed smartphone reader. The smartphone-read LFA signals for positive clinical samples tested ( N = 15, with known Ct) were statistically different ( p < 0.001) from signals for negative clinical samples ( N = 11). The phage LFA technology combined with smartphone chemiluminescence imaging can enable the timely development of ultrasensitive, affordable point-of-care testing platforms for SARS-CoV-2 and beyond. 

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4. A novel technology for home monitoring of lupus nephritis that tracks the pathogenic urine biomarker ALCAM

   https://doi.org/10.3389/fimmu.2022.1044743  

   Lei, Rongwei; Vu, Binh; Kourentzi, Katerina; Soomro, Sanam; Danthanarayana, Adheesha N.; Brgoch, Jakoah; Nadimpalli, Suma; Petri, Michelle; Mohan, Chandra; Willson, Richard C. (December 2022, Frontiers in Immunology)

   Introduction The gold standard for diagnosis of active lupus nephritis (ALN), a kidney biopsy, is invasive with attendant morbidity and cannot be serially repeated. Urinary ALCAM (uALCAM) has shown high diagnostic accuracy for renal pathology activity in ALN patients. Methods Lateral flow assays (LFA) for assaying uALCAM were engineered using persistent luminescent nanoparticles, read by a smartphone. The stability and reproducibility of the assembled LFA strips and freeze-dried conjugated nanoparticles were verified, as was analyte specificity. Results The LFA tests for both un-normalized uALCAM (AUC=0.93) and urine normalizer (HVEM)-normalized uALCAM (AUC=0.91) exhibited excellent accuracies in distinguishing ALN from healthy controls. The accuracies for distinguishing ALN from all other lupus patients were 0.86 and 0.74, respectively. Conclusion Periodic monitoring of uALCAM using this easy-to-use LFA test by the patient at home could potentially accelerate early detection of renal involvement or disease flares in lupus patients, and hence reduce morbidity and mortality. 

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5. Biomimetic Transparent Nanoplasmonic Meshes by Reverse‐Nanoimprinting for Bio‐Interfaced Spatiotemporal Multimodal SERS Bioanalysis

   https://doi.org/10.1002/smll.202204517  

   Garg, Aditya; Mejia, Elieser; Nam, Wonil; Vikesland, Peter; Zhou, Wei (September 2022, Small)

   Abstract Multicellular systems, such as microbial biofilms and cancerous tumors, feature complex biological activities coordinated by cellular interactions mediated via different signaling and regulatory pathways, which are intrinsically heterogeneous, dynamic, and adaptive. However, due to their invasiveness or their inability to interface with native cellular networks, standard bioanalysis methods do not allow in situ spatiotemporal biochemical monitoring of multicellular systems to capture holistic spatiotemporal pictures of systems‐level biology. Here, a high‐throughput reverse nanoimprint lithography approach is reported to create biomimetic transparent nanoplasmonic microporous mesh (BTNMM) devices with ultrathin flexible microporous structures for spatiotemporal multimodal surface‐enhanced Raman spectroscopy (SERS) measurements at the bio‐interface. It is demonstrated that BTNMMs, supporting uniform and ultrasensitive SERS hotspots, can simultaneously enable spatiotemporal multimodal SERS measurements for targeted pH sensing and non‐targeted molecular detection to resolve the diffusion dynamics for pH, adenine, and Rhodamine 6G molecules in agarose gel. Moreover, it is demonstrated that BTNMMs can act as multifunctional bio‐interfaced SERS sensors to conduct in situ spatiotemporal pH mapping and molecular profiling ofEscherichia colibiofilms. It is envisioned that the ultrasensitive multimodal SERS capability, transport permeability, and biomechanical compatibility of the BTNMMs can open exciting avenues for bio‐interfaced multifunctional sensing applications both in vitro and in vivo. 

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