Development Of Point-of-care Testing Research Articles

A Nanobody/Monoclonal Antibody "hybrid" sandwich technology offers an improved immunoassay strategy for detection of African trypanosome infections.

The scarcity of reliable devices for diagnosis of Animal African trypanosomiasis (AAT) presents a limitation to control of the disease. Existing high-sensitivity technologies such as PCR are costly, laborious, time-consuming, complex, and require skilled personnel. Hence, utilisation of most diagnostics for AAT is impracticable in rural areas, where the disease occurs. A more accessible point-of-care test (POCT) capable of detecting cryptic active infection, without relying on expensive equipment, would facilitate AAT detection. In turn, early management, would reduce disease incidence and severity. Today, several ongoing research projects aim at modifying complex immunoassays into POCTs. In this context, we report the development of an antigen (Ag) detection sandwich ELISA prototype for diagnosis of T. congolense infections, which is comprised of nanobody (Nb) and monoclonal antibody (mAb) reagents. The Nb474H used here, originated from a past study. Briefly, the Nb was engineered starting from mRNA of peripheral blood lymphocytes of an alpaca immunized with soluble lysate of Trypanosoma congolense (TC13). T. congolense glycosomal fructose-1,6-bisphosphate aldolase (TcoALD) was discovered as the cognate Ag of Nb474H. In this study, splenocytes were harvested from a mouse immunized with recombinant TcoALD and fused with NS01 cells to generate a hybridoma library. Random screening of the library on TcoALD retrieved a lone binder, designated IgM8A2. Using Nb474H as Ag-capture reagent in combination with the IgM8A2 monoclonal antibody Ag-detection reagent resulted in a tool that effectively detects native TcoALD released during infection by T. congolense parasites. Hitherto, development of POCT for detection of active trypanosome infection is elusive. The Nanobody/Monoclonal Antibody (Nb/mAb) "hybrid" sandwich technology offers prospects for exploration, using the unique specificity of Nb as a key determinant in Ag capturing, while using the versatility of monoclonal Ab to adapt to various detection conditions.

A laser-induced graphene-based electrochemical immunosensor for nucleic acid methylation detection.

The detection of methylation in DNA and RNA is essential for the diagnosis and treatment of a wide range of diseases. A one-step fabricated laser-induced graphene (LIG) electrode has received increasing attention due to its good electrical conductivity, large specific surface area, ease of miniaturization, low cost and flexibility. Herein, a potential biosensor for N6-methyladenosine (m6A-RNA) and 5-methylcystosine-single strand DNA (5mC-ssDNA) detection was designed. The aim of this paper is to address the problem of detecting the m6A-RNA and 5mC-ssDNA content in cells. By stepwise modification of gold nanoparticles (AuNPs), sulfhydryl-modified nucleic acid chains, biotin-modified antibodies, and streptavidin-modified horseradish peroxidase (SA-HRP) at the LIG electrode, the peak current responses exhibited an increase proportional to the concentration of m6A-RNA and 5mC-ssDNA in the hydrogen peroxide-hydroquinone (H2O2-HQ) system. This method demonstrated a low detection limit of 2.81 pM for m6A-RNA and 9.53 pM for 5mC-ssDNA, with a linear detection range of 0.01 nM to 10 nM for both targets. The regression equation was determined as ΔI = 4.83 log c + 12.32 (R2 = 0.9980) for m6A-RNA and ΔI = 9.82 log c + 22.09 (R2 = 0.9903) for 5mC-ssDNA. Our method has good selectivity toward different detection targets of nucleic acid chains, stability for long-term storage and consecutive scanning (RSD of 9.42% and 2.08%, respectively) and reproducibility of 5 electrodes (RSD of 6.85%). This method utilizes gold-sulfur bonding to immobilize the detection target, which improves the conductivity of the LIG electrode and introduces an amplified portion of the signal by taking advantage of antigen-antibody specific binding. Thus, dual detection of m6A-RNA and 5mC-ssDNA was realized. Importantly, this approach is successfully applied for the detection of targets in spiked samples extracted from HeLa cells, suggesting its potential for clinical applications and providing a new perspective for the development of point-of care testing (POCT) techniques.

Towards the development of cost-effective point-of-care diagnostic tools for poverty-related infectious diseases in sub-Saharan Africa.

In this review, we examine the current landscape of point-of-care testing (POCT) diagnostic tools designed for poverty-related infectious diseases (PRIDs) in sub-Saharan Africa (sSA) while delineating key avenues for future advancements. Our analysis encompasses both established and emerging diagnostic methods for PRIDs, addressing the persistent challenges in POCT tool development and deployment, such as cost, accessibility, and reliability. We emphasize recent advancements in POCT diagnostic tools as well as platforms poised to enhance diagnostic testing in sSA. Recognizing the urgency for affordable and widely accessible POCT diagnostic tools to detect PRIDs in sSA, we advocate for a multidisciplinary approach. This approach integrates current and emerging diagnostic methods, explicitly addressing challenges hindering point-of-care (POC) tool development. Furthermore, it recognizes the profound impact of misdiagnosis on public and global health, emphasizing the need for effective tools. To facilitate the successful development and implementation of POCT diagnostic tools in sSA, we propose strategies including the creation of multi-analyte detection POCT tools, the implementation of education and training programs, community engagement initiatives, fostering public-private collaborations, and the establishment of reliable supply chains. Through these concerted efforts, we aim to accelerate the development of POCT in the sSA region, ensuring its effectiveness and accessibility in addressing the diagnostic challenges associated with PRIDs.

Coupling hCG-based protease sensors with a commercial pregnancy test strip for simple analyses of protease activities

Proteases play an essential role in many cellular processes, and consequently, abnormalities in their activities are related to various diseases. Methods have been developed to measure the activity of these enzymes, but most involve sophisticated instruments or complicated procedures, which hampers the development of a point-of-care test (POCT). Here, we propose a strategy for developing simple and sensitive methods to analyze protease activity using commercial pregnancy test strips that detect human chorionic gonadotropin (hCG). hCG was engineered to have site-specific conjugated biotin and a peptide sequence, which can be cleaved by a target protease, between hCG and biotin. hCG protein was immobilized on streptavidin-coated beads, resulting in a protease sensor. The hCG-immobilized beads were too large to flow through the membrane of the hCG test strip and yielded only one band in the control line. When the peptide linker was hydrolyzed by the target protease, hCG was released from the beads, and the signal appeared in both the control and test lines. Three protease sensors for matrix metalloproteinase-2, caspase-3, and thrombin were constructed by replacing the protease-cleavable peptide linker. The combination of the protease sensors and a commercial pregnancy strip enabled the specific detection of each protease in the picomolar range, with a 30-min incubation of the hCG-immobilized beads and samples. The modular design of the protease sensor and simple assay procedure will facilitate the development of POCTs for various protease disease markers.

Paper-based colorimetric sensors for point-of-care testing.

By eliminating the need for sample transportation and centralized laboratory analysis, point-of-care testing (POCT) enables on-the-spot testing, with results available within minutes, leading to improved patient management and overall healthcare efficiency. Motivated by the rapid development of POCT, paper-based colorimetric sensing, a powerful analytical technique that exploits the changes in color or absorbance of a chemical species to detect and quantify analytes of interest, has garnered increasing attention. In this review, we strive to provide a bird's eye view of the development landscape of paper-based colorimetric sensors that harness the unique properties of paper to create low-cost, easy-to-use, and disposable analytical devices, thematically covering both fundamental aspects and categorized applications. In the end, we authors summarized the review with the remaining challenges and emerging opportunities. Hopefully, this review will ignite new research endeavors in the realm of paper-based colorimetric sensors.

Application of Janus Particles in Point-of-Care Testing.

Janus particles (JPs), named after the two-faced Roman god, are asymmetric particles with different chemical properties or polarities. JPs have been widely used in the biomedical field in recent years, including as drug carriers for targeted controlled drug release and as biosensors for biological imaging and biomarker detection, which is crucial in the early detection and treatment of diseases. In this review, we highlight the most recent advancements made with regard to Janus particles in point-of-care testing (POCT). Firstly, we introduce several commonly used methods for preparing Janus particles. Secondly, we present biomarker detection using JPs based on various detection methods to achieve the goal of POCT. Finally, we discuss the challenges and opportunities for developing Janus particles in POCT. This review will facilitate the development of POCT biosensing devices based on the unique properties of Janus particles.

Plasmonic Approaches for the Detection of SARS-CoV-2 Viral Particles.

The ongoing highly contagious Coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), underlines the fundamental position of diagnostic testing in outbreak control by allowing a distinction of the infected from the non-infected people. Diagnosis of COVID-19 remains largely based on reverse transcription PCR (RT-PCR), identifying the genetic material of the virus. Molecular testing approaches have been largely proposed in addition to infectivity testing of patients via sensing the presence of viral particles of SARS-CoV-2 specific structural proteins, such as the spike glycoproteins (S1, S2) and the nucleocapsid (N) protein. While the S1 protein remains the main target for neutralizing antibody treatment upon infection and the focus of vaccine and therapeutic design, it has also become a major target for the development of point-of care testing (POCT) devices. This review will focus on the possibility of surface plasmon resonance (SPR)-based sensing platforms to convert the receptor-binding event of SARS-CoV-2 viral particles into measurable signals. The state-of-the-art SPR-based SARS-CoV-2 sensing devices will be provided, and highlights about the applicability of plasmonic sensors as POCT for virus particle as well as viral protein sensing will be discussed.

Research Progress and Future Trends of Microfluidic Paper-Based Analytical Devices in In-Vitro Diagnosis.

In vitro diagnosis (IVD) has become a hot topic in laboratory research and achievement transformation. However, due to the high cost, and time-consuming and complex operation of traditional technologies, some new technologies are being introduced into IVD, to solve the existing problems. As a result, IVD has begun to develop toward point-of-care testing (POCT), a subdivision field of IVD. The pandemic has made governments and health institutions realize the urgency of accelerating the development of POCT. Microfluidic paper-based analytical devices (μPADs), a low-cost, high-efficiency, and easy-to-operate detection platform, have played a significant role in advancing the development of IVD. μPADs are composed of paper as the core material, certain unique substances as reagents for processing the paper, and sensing devices, as auxiliary equipment. The published reviews on the same topic lack a comprehensive and systematic introduction to μPAD classification and research progress in IVD segmentation. In this paper, we first briefly introduce the origin of μPADs and their role in promoting IVD, in the introduction section. Then, processing and detection methods for μPADs are summarized, and the innovative achievements of μPADs in IVD are reviewed. Finally, we discuss and prospect the upgrade and improvement directions of μPADs, in terms of portability, sensitivity, and automation, to help researchers clarify the progress and overcome the difficulties in subsequent μPAD research.

Recent Advances and Applications in Paper-Based Devices for Point-of-Care Testing.

Point-of-care testing (POCT), as a portable and user-friendly technology, can obtain accurate test results immediately at the sampling point. Nowadays, microfluidic paper-based analysis devices (μPads) have attracted the eye of the public and accelerated the development of POCT. A variety of detection methods are combined with μPads to realize precise, rapid and sensitive POCT. This article mainly introduced the development of electrochemistry and optical detection methods on μPads for POCT and their applications on disease analysis, environmental monitoring and food control in the past 5 years. Finally, the challenges and future development prospects of μPads for POCT were discussed.

2020 STD Prevention Conference: Disrupting Epidemics and Dismantling Disparities in the Time of COVID-19.

2020 STD Prevention Conference: Disrupting Epidemics and Dismantling Disparities in the Time of COVID-19.

Cardiac biomarker measurement by point of care testing – Development, rationale, current state and future developments

Cardiac biomarker measurements are integral to the diagnosis and management of patients presenting with breathlessness and chest pain. Measurement of B type natriuretic peptide either directly or of the N-terminal portion of the prohormone although possible by point of care testing (POCT) has largely become a laboratory test. Measurement of the cardiac troponins cardiac troponin T (cTnT) and cardiac troponin I (cTnI) can easily and accurately be performed by POCT. The situation has been complicated by the development of high sensitivity assays for cTnT and cTnI and the subsequent development of rapid rule out algorithms allowing patient categorisation and discharge on admission and 1 to 2 h following admission. This article reviews the development of POCT for cardiac biomarkers, the evidence base comparing POCT with central laboratory testing, its strengths and limitations, and how POCT fits into the world of high sensitivity troponin assays. It also discusses what evidence there is that POCT can form part of rapid decision-making strategies and how this applies in an era of algorithms based on and is derived from measurement of high sensitivity troponin in the central laboratory.

Importance and Feasibility of Point-of-care Testing in Takayasu’s Arteritis

Takayasu’s arteritis (TAK) is known to be a unique, rare, and chronic vasculitis disease that affects large elastic arteries such as aorta and its major branches. TAK is characterized by adventitial thickening, weak pulses and ocular disturbances. The prognosis and diagnosis of TAK are challenging due to the non-specific, silent, or paucisymptomatic presentation of the disease. The effective understanding of TAK lies with the timely recognition of the symptoms and a rapid diagnosis of the disease. Point-of-care testing (POCT) is vital for the quick and reliable detection of parameters near bedside for disease diagnosis, assessment, monitoring, and therapeutics management. Ultrasonography is the most reliable POCT technique. Moreover, erythrocyte sedimentation rate and C-reactive protein assay are the two most valuable non-imaging POCT tests used to determine inflammation and onset of the disease. Other potential biomarkers such as matrix metalloproteinases, soluble receptor for advanced glycation end products, interleukin (IL)-6, and IL-18 have also been advocated for tracking the progression of TAK. Furthermore, vasculitis associated-anti-neutrophil cytoplasmic antibodies have also been reported to reflect the inflammatory phase of the disease. Therefore, the development of POCT based on these blood-based biomarkers may help in quick clinical decision-making for early diagnosis of TAK and targeted therapeutics to improve clinical outcome in patients suffering from this debilitating disease.

Point-of-care testing in the diagnosis of infectious diseases

Rapid diagnosis is important for the prevention and control of infectious disease. Point-of-care testing (POCT) technology is simple, mobile, rapid, sensitive and accurate. In recent years, POCT has been widely used in the detection of infectious agents. This article reviews the development of POCT in the diagnosis of infectious diseases, focusing on immuno-chromatographic technology, microfluidic chip technology and loop-mediated isothermal amplification (LAMP) technology. Key words: Infections diseases; Point-of-care testing; Immunochromatography; Lab-on-A-chip devices; Nucleic acid amplification techniques

Renal point-of-care testing: collaboration between biomedical scientists and community pharmacists

ABSTRACTThe incidence of chronic kidney disease (CKD) is increasing with the prevalence of obesity, diabetes mellitus, elevated blood pressure and other cardiovascular-renal risk factors. Wider point-of-care testing (POCT) strategies in the community setting are needed to prevent CKD and delay progressive loss of renal function. Since publication of the Institute of Biomedical Science (IBMS) Point of Care Testing (Near-Patient Testing) Guidance on the Involvement of the Clinical Laboratory, the biomedical scientist can now provide enhanced services including tests for blood glucose, cholesterol, and monitor blood pressure. Under the current pharmacy contract, community pharmacies are now often equipped with sophisticated record access systems and interfaces to monitor drug interactions in suspect/high-risk CKD patients. Current facilities can be utilised further allowing pharmacists more clinical involvement based on community need. Further development of POCT in the community involving collaborations between biomedical scientists and community pharmacists would allow wider service availability in primary care and might be advantageous in suspect/high-risk patients.

Rapid Diagnosis of Sepsis

Unacceptable mortality rates and excessively high costs of sepsis can be attributed in part to delayed pathogen identification, poorly focused treatment, and unjustified antimicrobial use. Rapid diagnosis of pathogens, immediately focused antibiotics, and quick interruption of dysfunctional sepsis cascades could optimize clinical value. Value analysis constructs evidence-based hypotheses, called value propositions, that form a value model for the use of rapid diagnosis and point-of-care testing (POCT) in sepsis. Rapid diagnostic methods, such as nucleic acid testing (NAT), can assure fast therapeutic turnaround time. The authors applied NAT hypothetically to different scenarios for which they performed value analysis of patient and hospital costs and of medical outcomes. Value propositions were constructed for both adult and pediatric clinical situations. The authors conclude that rapid diagnosis by NAT can reduce the probability of complications, allow physicians to immediately focus treatment, and decrease both costs and mortality. Expected benefits for both pediatric and adult patients validate the need for continued NAT research and development of POCT for critically ill patients with sepsis.