Design and implementation of a digital site-less clinical study of serial rapid antigen testing to identify asymptomatic SARS-CoV-2 infection.

Background: Rapid antigen detection tests (Ag-RDT) for SARS-CoV-2 with emergency use authorization generally include a condition of authorization to evaluate the test's performance in asymptomatic individuals when used serially. We aim to describe a novel study design that was used to generate regulatory-quality data to evaluate the serial use of Ag-RDT in detecting SARS-CoV-2 virus among asymptomatic individuals. Methods: This prospective cohort study used a siteless, digital approach to assess longitudinal performance of Ag-RDT. Individuals over 2 years old from across the USA with no reported COVID-19 symptoms in the 14 days prior to study enrollment were eligible to enroll in this study. Participants throughout the mainland USA were enrolled through a digital platform between October 18, 2021 and February 15, 2022. Participants were asked to test using Ag-RDT and molecular comparators every 48 hours for 15 days. Enrollment demographics, geographic distribution, and SARS-CoV-2 infection rates are reported. Key Results: A total of 7361 participants enrolled in the study, and 492 participants tested positive for SARS-CoV-2, including 154 who were asymptomatic and tested negative to start the study. This exceeded the initial enrollment goals of 60 positive participants. We enrolled participants from 44 US states, and geographic distribution of participants shifted in accordance with the changing COVID-19 prevalence nationwide. Conclusions: The digital site-less approach employed in the "Test Us At Home" study enabled rapid, efficient, and rigorous evaluation of rapid diagnostics for COVID-19 and can be adapted across research disciplines to optimize study enrollment and accessibility.

Low-Cost, Real-Time Polymerase Chain Reaction System with Integrated RNA Extraction.

Rapid, easy-to-use, and low-cost systems for biological sample testing are important for point-of-care diagnostics and various other health applications. The recent pandemic of Coronavirus Disease 2019 (COVID-19) caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) showed an urgent need to rapidly and accurately identify the genetic material of SARS-CoV-2, an enveloped ribonucleic acid (RNA) virus, in upper respiratory specimens from people. In general, sensitive testing methods require genetic material extraction from the specimen. Unfortunately, current commercially available extraction kits are expensive and involve time-consuming and laborious extraction procedures. To overcome the difficulties associated with common extraction methods, we propose a simple enzymatic assay for the nucleic acid extraction step using heat mediation to improve the polymerase chain reaction (PCR) reaction sensitivity. Our protocol was tested on Human Coronavirus 229E (HCoV-229E) as an example, which comes from the large coronaviridae family of viruses that affect birds, amphibians, and mammals, of which SARS-CoV-2 is a member. The proposed assay was performed using a low-cost, custom-made, real-time PCR system that incorporates thermal cycling and fluorescence detection. It had fully customizable reaction settings to allow versatile biological sample testing for various applications, including point-of-care medical diagnosis, food and water quality testing, and emergency health situations. Our results show that heat-mediated RNA extraction is a viable extraction method when compared to commercial extraction kits. Further, our study showed that extraction has a direct impact on purified laboratory samples of HCoV-229E, but no direct impact on infected human cells. This is clinically relevant, as it allows us to circumvent the extraction step on clinical samples when using PCR.

Engineering highly thermostable Cas12b via de novo structural analyses for one-pot detection of nucleic acids.

CRISPR-Cas-based diagnostics have the potential to elevate nucleic acid detection. CRISPR-Cas systems can be combined with a pre-amplification step in a one-pot reaction to simplify the workflow and reduce carryover contamination. Here, we report an engineered Cas12b with improved thermostability that falls within the optimal temperature range (60°C-65°C) of reverse transcription-loop-mediated isothermal amplification (RT-LAMP). Using de novo structural analyses, we introduce mutations to wild-type BrCas12b to tighten its hydrophobic cores, thereby enhancing thermostability. The one-pot detection assay utilizing the engineered BrCas12b, called SPLENDID (single-pot LAMP-mediated engineered BrCas12b for nucleic acid detection of infectious diseases), exhibits robust trans-cleavage activity up to 67°C in a one-pot setting. We validate SPLENDID clinically in 80 serum samples for hepatitis C virus (HCV) and 66 saliva samples for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with high specificity and accuracy. We obtain results in as little as 20 min, and with the extraction process, the entire assay can be performed within an hour.

A handheld plug-and-play microfluidic liquid handling automation platform for immunoassays.

Lab-on-a-chip technologies and microfluidics have pushed miniaturized liquid handling to unprecedented precision, integration, and automation, which improved the reaction efficiency of immunoassays. However, most microfluidic immunoassay systems still require bulky infrastructures, such as external pressure sources, pneumatic systems, and complex manual tubing and interface connections. Such requirements prevent plug-and-play operation at the point-of-care (POC) settings. Here we present a fully automated handheld general microfluidic liquid handling automation platform with a plug-and-play 'clamshell-style' cartridge socket, a miniature electro-pneumatic controller, and injection-moldable plastic cartridges. The system achieved multi-reagent switching, metering, and timing control on the valveless cartridge using electro-pneumatic pressure control. As a demonstration, a SARS-CoV-2 spike antibody sandwich fluorescent immunoassay (FIA) liquid handling was performed on an acrylic cartridge without human intervention after sample introduction. A fluorescence microscope was used to analyze the result. The assay showed a limit of detection at 31.1 ng/mL, comparable to some previously reported enzyme-linked immunosorbent assays (ELISA). In addition to automated liquid handling on the cartridge, the system can operate as a 6-port pressure source for external microfluidic chips. A rechargeable battery with a 12 V 3000 mAh capacity can power the system for 42 h. The footprint of the system is 16.5 × 10.5 × 7 cm, and the weight is 801 g, including the battery. The system can find many other POC and research applications requiring complex liquid manipulation, such as molecular diagnostics, cell analysis, and on-demand biomanufacturing.

CoVSense: Ultrasensitive Nucleocapsid Antigen Immunosensor for Rapid Clinical Detection of Wildtype and Variant SARS-CoV-2.

The widespread accessibility of commercial/clinically-viable electrochemical diagnostic systems for rapid quantification of viral proteins demands translational/preclinical investigations. Here, Covid-Sense (CoVSense) antigen testing platform; an all-in-one electrochemical nano-immunosensor for sample-to-result, self-validated, and accurate quantification of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid (N)-proteins in clinical examinations is developed. The platform's sensing strips benefit from a highly-sensitive, nanostructured surface, created through the incorporation of carboxyl-functionalized graphene nanosheets, and poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) conductive polymers, enhancing the overall conductivity of the system. The nanoengineered surface chemistry allows for compatible direct assembly of bioreceptor molecules. CoVSense offers an inexpensive (<$2 kit) and fast/digital response (<10 min), measured using a customized hand-held reader (<$25), enabling data-driven outbreak management. The sensor shows 95% clinical sensitivity and 100% specificity (Ct<25), and overall sensitivity of 91% for combined symptomatic/asymptomatic cohort with wildtype SARS-CoV-2 or B.1.1.7 variant (N = 105, nasal/throat samples). The sensor correlates the N-protein levels to viral load, detecting high Ct values of ≈35, with no sample preparation steps, while outperforming the commercial rapid antigen tests. The current translational technology fills the gap in the workflow of rapid, point-of-care, and accurate diagnosis of COVID-19.

COVID-19 surveillance in wastewater: An epidemiological tool for the monitoring of SARS-CoV-2.

The coronavirus disease 2019 (COVID-19) pandemic has prompted a lot of questions globally regarding the range of information about the virus's possible routes of transmission, diagnostics, and therapeutic tools. Worldwide studies have pointed out the importance of monitoring and early surveillance techniques based on the identification of viral RNA in wastewater. These studies indicated the presence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in human feces, which is shed via excreta including mucus, feces, saliva, and sputum. Subsequently, they get dumped into wastewater, and their presence in wastewater provides a possibility of using it as a tool to help prevent and eradicate the virus. Its monitoring is still done in many regions worldwide and serves as an early "warning signal"; however, a lot of limitations of wastewater surveillance have also been identified.

Liposome-based high-throughput and point-of-care assays toward the quick, simple, and sensitive detection of neutralizing antibodies against SARS-CoV-2 in patient sera.

The emergence of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) in 2019 caused an increased interest in neutralizing antibody tests to determine the immune status of the population. Standard live-virus-based neutralization assays such as plaque-reduction assays or pseudovirus neutralization tests cannot be adapted to the point-of-care (POC). Accordingly, tests quantifying competitive binding inhibition of the angiotensin-converting enzyme 2 (ACE2) receptor to the receptor-binding domain (RBD) of SARS-CoV-2 by neutralizing antibodies have been developed. Here, we present a new platform using sulforhodamine B encapsulating liposomes decorated with RBD as foundation for the development of both a fluorescent, highly feasible high-throughput (HTS) and a POC-ready neutralizing antibody assay. RBD-conjugated liposomes are incubated with serum and subsequently immobilized in an ACE2-coated plate or mixed with biotinylated ACE2 and used in test strip with streptavidin test line, respectively. Polyclonal neutralizing human antibodies were shown to cause complete binding inhibition, while S309 and CR3022 human monoclonal antibodies only caused partial inhibition, proving the functionality of the assay. Both formats, the HTS and POC assay, were then tested using 20 sera containing varying titers of neutralizing antibodies, and a control panel of sera including prepandemic sera and reconvalescent sera from respiratory infections other than SARS-CoV-2. Both assays correlated well with a standard pseudovirus neutralization test (r = 0.847 for HTS and r = 0.614 for POC format). Furthermore, excellent correlation (r = 0.868) between HTS and POC formats was observed. The flexibility afforded by liposomes as signaling agents using different dyes and sizes can hence be utilized in the future for a broad range of multianalyte neutralizing antibody diagnostics.

Rapid point-of-care detection of SARS-CoV-2 RNA with smartphone-based upconversion luminescence diagnostics.

Accurate COVID-19 screening via molecular technologies is still hampered by bulky instrumentation, complicated procedure, high cost, lengthy testing time, and the need for specialized personnel. Herein, we develop point-of-care upconversion luminescence diagnostics (PULD), and a streamlined smartphone-based portable platform facilitated by a ready-to-use assay for rapid SARS-CoV-2 nucleocapsid (N) gene testing. With the complementary oligo-modified upconversion nanoprobes and gold nanoprobes specifically hybridized with the target N gene, the luminescence resonance energy transfer effect leads to a quenching of fluorescence intensity that can be detected by the easy-to-use diagnostic system. A remarkable detection limit of 11.46 fM is achieved in this diagnostic platform without the need of target amplification, demonstrating high sensitivity and signal-to-noise ratio of the assay. The capability of the developed PULD is further assessed by probing 9 RT-qPCR-validated SARS-CoV-2 variant clinical samples (B.1.1.529/Omicron) within 20 min, producing reliable diagnostic results consistent with those obtained from a standard fluorescence spectrometer. Importantly, PULD is capable of identifying the positive COVID-19 samples with superior sensitivity and specificity, making it a promising front-line tool for rapid, high-throughput screening and infection control of COVID-19 or other infectious diseases.

Mucormycosis diagnosis revisited: Current and emerging diagnostic methodologies for the invasive fungal infection (Review).

Mucormycosis, which is a life threatening condition, is one of the side effects experienced by post-COVID-19 patients. Early identification and timely treatment are essential to stop the dissemination of the disease, since invasive mucormycosis has a very high fatality rate and significant disease dispersion. Conventional diagnostic techniques, including clinical diagnosis, serology, histopathology and radiology, have limitations in diagnosing the disease at an early stage. This warrants the need for advanced diagnostic tools such as nucleic acid diagnostics, advanced serological tests (ELISpot), PCR (pan-Mucorale test) and multiplex PCR. These techniques have been introduced to identify this invasive fungal infection at an incipient stage, thereby helping clinicians to prevent adverse outcomes. The use of biosensors and micro-needle based diagnostic methodologies will pave the way for devising more point-of-care tests that can be employed for the detection of mucormycosis at an incipient stage. The present review discusses the current techniques available and their drawbacks, and the usefulness of advanced diagnostic tools. Furthermore, the possibility of using future diagnostic methods for the diagnosis of mucormycosis is highlighted.

A Nitrocellulose Paper-Based Multi-Well Plate for Point-of-Care ELISA.

Low-cost diagnostic tools for point-of-care immunoassays, such as the paper-based enzyme-linked immunoassay (ELISA), have become increasingly important, especially so in the recent COVID-19 pandemic. ELISA is the gold-standard antibody/antigen sensing method. This paper reports an easy-to-fabricate nitrocellulose (NC) paper plate, coupled with a desktop scanner for ELISA, which provides a higher protein immobilization efficiency than the conventional cellulose paper-based ELISA platforms. The experiments were performed using spiked samples for the direct ELISA of rabbit IgG with a limit of detection (LOD) of 1.016 µg/mL, in a measurement range of 10 ng/mL to 1 mg/mL, and for the sandwich ELISA of sperm protein (SP-10) with an LOD of 88.8 ng/mL, in a measurement range of 1 ng/mL to 100 µg/mL. The described fabrication method, based on laser-cutting, is a highly flexible one-step laser micromachining process, which enables the rapid production of low-cost NC paper-based multi-well plates with different sizes for the ELISA measurements.

Amplification Free Detection of SARS-CoV-2 Using Multi-Valent Binding.

We present the development of electrochemical impedance spectroscopy (EIS)-based biosensors for sensitive detection of SARS-CoV-2 RNA using multi-valent binding. By increasing the number of probe-target binding events per target molecule, multi-valent binding is a viable strategy for improving the biosensor performance. As EIS can provide sensitive and label-free measurements of nucleic acid targets during probe-target hybridization, we used multi-valent binding to build EIS biosensors for targeting SARS-CoV-2 RNA. For developing the biosensor, we explored two different approaches including probe combinations that individually bind in a single-valent fashion and the probes that bind in a multi-valent manner on their own. While we found excellent biosensor performance using probe combinations, we also discovered unexpected signal suppression. We explained the signal suppression theoretically using inter- and intra-probe hybridizations which confirmed our experimental findings. With our best probe combination, we achieved a LOD of 182 copies/µL (303 aM) of SARS-CoV-2 RNA and used these for successful evaluation of patient samples for COVID-19 diagnostics. We were also able to show the concept of multi-valent binding with shorter probes in the second approach. Here, a 13-nt-long probe has shown the best performance during SARS-CoV-2 RNA binding. Therefore, multi-valent binding approaches using EIS have high utility for direct detection of nucleic acid targets and for point-of-care diagnostics.

The Future of Point-of-Care Nucleic Acid Amplification Diagnostics after COVID-19: Time to Walk the Walk.

Since the onset of the COVID-19 pandemic, over 610 million cases have been diagnosed and it has caused over 6.5 million deaths worldwide. The crisis has forced the scientific community to develop tools for disease control and management at a pace never seen before. The control of the pandemic heavily relies in the use of fast and accurate diagnostics, that allow testing at a large scale. The gold standard diagnosis of viral infections is the RT-qPCR. Although it provides consistent and reliable results, it is hampered by its limited throughput and technical requirements. Here, we discuss the main approaches to rapid and point-of-care diagnostics based on RT-qPCR and isothermal amplification diagnostics. We describe the main COVID-19 molecular diagnostic tests approved for self-testing at home or for point-of-care testing and compare the available options. We define the influence of specimen selection and processing, the clinical validation, result readout improvement strategies, the combination with CRISPR-based detection and the diagnostic challenge posed by SARS-CoV-2 variants for different isothermal amplification techniques, with a particular focus on LAMP and recombinase polymerase amplification (RPA). Finally, we try to shed light on the effect the improvement in molecular diagnostics during the COVID-19 pandemic could have in the future of other infectious diseases.

Designing and Piloting of a Mobile Learning Curriculum for Quality Point-Of-Care Diagnostics Services in Rural Clinics of KwaZulu-Natal, South Africa.

Background: The use of mobile technology has been reported to help improve access to education for people in remote areas. However, there is limited evidence of its adoption in resource-limited settings. The aim of this study was to utilize stakeholder generated ideas to design and pilot a mobile learning curriculum, with the purpose of facilitating training to improve the quality of point-of-care diagnostics services in KwaZulu-Natal (KZN) rural clinics. Methods: Nominal Group Technique was employed to enable collaboration with stakeholders in designing and piloting of a POC diagnostics curriculum. Stakeholders were selected from 11 KZN districts to participate in a clinic-based piloting of the curriculum using an online application. The application was designed in collaboration with a teacher training institute in Durban. Moodle was used as an established reliable online learning management system. During piloting, quantitative and qualitative data were generated and analyzed using descriptive statistics and content analysis. Findings and Conclusion: Guided by the Nominal Group Technique results, five delivery modes for curriculum content through Mobile Learning were generated. An interactive course page was created on the Moodle site, titled: Quality HIV Point of Care Diagnostics Curriculum Delivery for Nurses in Rural Areas. The course content consisted of three teaching units, activities, an online quiz and an online survey. An analytic-algorithm built into the online course enabled monitoring of participation and assessment outcomes automatically. At piloting, 64% of the invited representative clinics were able to access the course, with 47% meeting the course completion requirements. All the participants achieved the set pass mark of 75% with an average of 87%. The activity completion report showed that topics presented through images, videos and simple text were accessed more than those presented as attachments of national documents. Despite poor network coverage and limited access to mobile technology, exacerbated by Covid-19 related restrictions, Point of care diagnostics Mobile Learning curriculum was well-received in participating rural clinics. Recommendations relating to course improvement and access, included extending collaboration with specialists in eHealth systems development and with South African cell phone network providers.

Recent Advances of Representative Optical Biosensors for Rapid and Sensitive Diagnostics of SARS-CoV-2.

The outbreak of Corona Virus Disease 2019 (COVID-19) has again emphasized the significance of developing rapid and highly sensitive testing tools for quickly identifying infected patients. Although the current reverse transcription polymerase chain reaction (RT-PCR) diagnostic techniques can satisfy the required sensitivity and specificity, the inherent disadvantages with time-consuming, sophisticated equipment and professional operators limit its application scopes. Compared with traditional detection techniques, optical biosensors based on nanomaterials/nanostructures have received much interest in the detection of SARS-CoV-2 due to the high sensitivity, high accuracy, and fast response. In this review, the research progress on optical biosensors in SARS-CoV-2 diagnosis, including fluorescence biosensors, colorimetric biosensors, Surface Enhancement Raman Scattering (SERS) biosensors, and Surface Plasmon Resonance (SPR) biosensors, was comprehensively summarized. Further, promising strategies to improve optical biosensors are also explained. Optical biosensors can not only realize the rapid detection of SARS-CoV-2 but also be applied to judge the infectiousness of the virus and guide the choice of SARS-CoV-2 vaccines, showing enormous potential to become point-of-care detection tools for the timely control of the pandemic.

Rapid and ultrasensitive detection of SARS-CoV-2 spike protein based on upconversion luminescence biosensor for COVID-19 point-of-care diagnostics.

Here, we firstly introduce a detection system consisting of upconversion nanoparticles (UCNPs) and Au nanorods (AuNRs) for an ultrasensitive, rapid, quantitative and on-site detection of SARS-CoV-2 spike (S) protein based on Förster resonance energy transfer (FRET) effect. Briefly, the UCNPs capture the S protein of lysed SARS-CoV-2 in the swabs and subsequently they are bound with the anti-S antibodies modified AuNRs, resulting in significant nonradiative transitions from UCNPs (donors) to AuNRs (acceptors) at 480 nm and 800 nm, respectively. Notably, the specific recognition and quantitation of S protein can be realized in minutes at 800 nm because of the low autofluorescence and high Yb-Tm energy transfer in upconversion process. Inspiringly, the limit of detection (LOD) of the S protein can reach down to 1.06 fg mL-1, while the recognition of nucleocapsid protein is also comparable with a commercial test kit in a shorter time (only 5 min). The established strategy is technically superior to those reported point-of-care biosensors in terms of detection time, cost, and sensitivity, which paves a new avenue for future on-site rapid viral screening and point-of-care diagnostics.

MICROFLUIDIC DIAGNOSTIC AND POINT OF CARE DETECTION SYSTEM FOR SARS-COV-2 FROM SALIVA

We present a nucleic acid-based point-of-care diagnostic for the detection of the SARS-CoV-2 from saliva using an additively manufactured microfluidic cartridge. The assay uses reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) to detect the presence of SARS-CoV-2 RNA on-cartridge in a point-of-care optical detection system based on a smartphone. We show positive results within the 10-30 minutes range and integrated biological controls on the cartridge. We demonstrate the microfluidic diagnostic with human patient samples, with results that are consistent with the off-cartridge validation. © 2021 MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. All rights reserved.

Multiplexed Point-of-Care Electrical Detection of COVID-19 Biomarkers Using Enzymatically Amplified Metallization on Nanostructured Surfaces

We present a multiplexed, electronic enzyme-linked immunosorbent assay (E2LISA) microchip for direct electrical detection and quantitation of multiple biomarkers from a single microliter-scale drop of sample. Spatially distinct spots on the microchip, each containing an interdigitated microelectrode array, are coated with specific capture agents and used to bind different analytes. Enzyme-labeled probes are then used to convert this analyte binding to an electrical impedance signal via the amplified, localized deposition of silver on the nanostructured, catalytic surface of the chip prepared using gold nanoparticles. We use this microchip with a custom handheld, cellphone interfaced reader to detect COVID-19 biomarkers including antigen-specific antibodies and viral antigens. Further, we demonstrate the multiplexed measurement of distinct antibody responses in serum samples from convalescent COVID-19 patients versus uninfected vaccine recipients. © 2021 MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. All rights reserved.

Rapid SARS-CoV-2 testing with duplexed recombinase polymerase amplification and a bacteriophage internal control

Without global mass vaccination, COVID-19 will continue to infect and cause serious illness, disproportionately in low- and middle-income countries. Point-of-care and home-based nucleic acid amplification tests (NAATs) are valuable tools to control COVID-19 transmission. Here we present a rapid isothermal NAAT for duplexed detection of SARS-CoV-2 and an MS2 bacteriophage internal control. This assay amplifies RNA in less than 15 minutes, utilizes a low temperature of 39°C, and has fluorescence or visual lateral flow readout. This positions our assay for use in low-cost paper-based nucleic acid diagnostic devices for ultrasensitive and reliable COVID-19 detection in POC or home-based settings. © 2021 MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. All rights reserved.

HANDHELD AND ULTRAFAST PHOTOTHERMAL QPCR SYSTEM FOR RAPID DETECTION OF SARS-COV-2

Rapid polymerase chain reaction (PCR) utilizing plasmon-driven photothermal cycling requires real-time quantification of amplicons during PCR and miniaturization of real-time PCR (qPCR) system for point-of-care (POC) diagnostics. In this work, we have demonstrated handheld photothermal qPCR system with disposable aluminum PCR chips for the ultrafast amplification and real-time quantification of plasmids expressing SARSCoV-2 envelope protein within 5 min. This novel system provides stable and useful point-of-care diagnostic platform for prevention of fast-spreading pandemic in airport and harbor. © 2021 MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. All rights reserved.

Laser-induced breakdown spectroscopy (LIBS)-based assay for pointof-care (POC) detection of cytokines in COVID-19 infection

Accumulating evidence suggests that cytokine storm syndrome (CSS) induced by the SARS-CoV-2 may be the ultimate cause of acute respiratory distress syndrome (ARDS), resulting in severe outcomes of COVID- 19 infection and potentially death. Elevated levels of serum interleukin 6 (IL-6) correlate with the occurrence of respiratory failure, ARDS, and adverse clinical outcomes in many COVID-19 patients. The currently available clinical cytokine tests are costly, time-consuming, and require skilled technicians to execute. There is an unmet need for rapid, affordable, robust, and sensitive tests for cytokine levels. Therefore, this study aimed to develop a cost-effective system for quantitative detection of cytokines that can be used in the point-of-care (POC) format within a few minutes of blood collection. Our approach combines detection based on laser-induced breakdown spectroscopy with a lateral flow immunoassay (LIBS- LFIA) to deliver a quantitative clinical analysis platform with multiplexing capability. Lanthanide-complexed polymers (LCPs) were selected as the labels to provide optimal quantitative performance when sensing signals from the test lines of LFIAs. For a prototype implementation and a proof-of-concept, we targeted IL-6 as it is one of the most critical pro-inflammatory cytokines. Our initial LIBS-LFIA biosensor achieved a limit of detection ( LOD) of 0.2298 mu g/mL of IL-6 within 15 minutes and further sensitivity increase is possible with optimization. Regardless, since high levels of IL-6 are reported for patients in crisis, this is more than adequate to identify patients with highly elevated cytokine levels. Our research provides evidence that rapid and accurate detection of cytokines for clinical diagnosis and prognosis of COVID-19 and other pathogenic infections using LIBS is highly feasible and compatible with the POC format.