Point Of Care Diagnostic Tool Research Articles

Lung ultrasound in respiratory therapy: a global reflective survey.

Lung ultrasound (LUS) is a non-invasive point of care diagnostic tool used to assess the presence and severity of various lung disorders for more than two decades. Within the healthcare professionals are the respiratory therapists (RTs) who play a vital role in managing ventilated and other patients requiring respiratory support, but the incorporation of LUS into their scope of practice has not been well highlighted. This international cross-sectional survey was specifically designed to evaluate the knowledge, attitude, and practice of RTs with respect to LUS. This observational cross-sectional study was conducted among RTs from different parts of the world using a questionnaire-based study tool. 514 RTs responded to all the questions and were considered for statistical analysis. Descriptive statistics, Analysis of variance, Fisher's exact, Chi-square, Bonferroni post-hoc analysis, and Binomial logistic regression analyses were performed to identify the significance of the data. From 22 countries, 514 RTs responded to the survey, with the major share from the middle eastern countries. Out of the 514 responders, 44.9% of the responders were in the age group of 23-30 years; 67.1% were bachelor's degree holders; and 40.9% of participants had more than 10 years of experience. The knowledge-based questions revealed that RTs with higher experience and academic qualification provided more positive responses. While in the attitude-related domain it is observed that standardized training in LUS helps them to enhance the current practice and to add LUS to the academic curriculum of respiratory therapy schools; however there remains barriers to practice LUS based on their responses. The practice-based questions revealed that RTs expect some additional seminars/workshops/webinars to be done on LUS frequently. More than half of the participants are found to be knowledgeable with a positive attitude and working towards the inclusion of LUS in respiratory therapy profession. It is concluded that respiratory therapists have a positive attribute towards the inclusion of lung ultrasound in their clinical practice. Providing more structured training for professional respiratory therapists and including lung ultrasound modules in the respiratory therapy school curriculum may facilitate mastering their diagnostic skills, thereby expanding the scope of practice.

Review of Gold Nanoparticles in Surface Plasmon-Coupled Emission Technology: Effect of Shape, Hollow Nanostructures, Nano-Assembly, Metal-Dielectric and Heterometallic Nanohybrids.

Point-of-care (POC) diagnostic platforms are globally employed in modern smart technologies to detect events or changes in the analyte concentration and provide qualitative and quantitative information in biosensing. Surface plasmon-coupled emission (SPCE) technology has emerged as an effective POC diagnostic tool for developing robust biosensing frameworks. The simplicity, robustness and relevance of the technology has attracted researchers in physical, chemical and biological milieu on account of its unique attributes such as high specificity, sensitivity, low background noise, highly polarized, sharply directional, excellent spectral resolution capabilities. In the past decade, numerous nano-fabrication methods have been developed for augmenting the performance of the conventional SPCE technology. Among them the utility of plasmonic gold nanoparticles (AuNPs) has enabled the demonstration of plethora of reliable biosensing platforms. Here, we review the nano-engineering and biosensing applications of AuNPs based on the shape, hollow morphology, metal-dielectric, nano-assembly and heterometallic nanohybrids under optical as well as biosensing competencies. The current review emphasizes the recent past and evaluates the latest advancements in the field to comprehend the futuristic scope and perspectives of exploiting Au nano-antennas for plasmonic hotspot generation in SPCE technology.

Kaedah Diagnostik Semasa dan Penggunaan Ujian Titik Penjagaan Pantas (POC) Bagi Mendiagnos Hiperkolesterolemia Famili (FH)

Familial hypercholesterolemia (FH) is an autosomal dominant inherited genetic disease characterized by increased concentrations of low-density lipoprotein (LDL-C) cholesterol in the blood. The risk of premature coronary heart disease in FH patients may increase without early treatment. Advancement in molecular biology techniques has enable early detection and diagnosis of FH. These techniques are cost-effective and have a shorter turnaround time. The current diagnostic tools available for FH diagnosis involving algorithm-based scoring criteria and various molecular diagnosis methods including next-generation sequencing (NGS), Sanger sequencing, Multiplex ligation-dependent probe amplification (MLPA) and DNA hybridisation assay are discussed in this review. However, molecular genetic testing is not widely available due to time-consuming procedures, high cost and requires trained personnel. Thus, this 36 review highlights the use of point of care (POC) testing as an approach to diagnose FH, particularly in countries lacking infrastructure and expertise in this field. Lateral flow testing (LFA) has gained attention as a POC diagnostic tool due to its simplicity, low cost and involved simple procedure and settings. The advantages of LFA made this technique a potential tool in addressing challenges in diagnosing FH, particularly for early diagnosis of family members.

Paper disc interfaced Prussian blue nanocube modified immunodevice for electrochemical detection of diverse biomarker at point of care

The detection of specific biomarkers is used in various phases of the diagnosis of plant and human diseases, from prognosis to monitoring. Herein, we report a Prussian blue nanocube-modified immunodevice interfaced with a paper disc for the detection of plant biomarkers via streptavidin-biotin recognition. The detection ability of the immunodevice was assessed using Potato virus X as a model biomarker and analyzed using cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy. The immunodevice displayed excellent performance for Potato virus X detection with a detection limit of 0.92 nM (3S/N). The selectivity of the fabricated Potato virus X immunodevice was investigated using closely associated antigens, such as potato aucuba mosaic virus, Potato virus Y, and Potato virus A. The Potato virus X immunodevice exhibited ∼ 90 % recovery in spiked complex plant samples with a relative error of ∼ 9 %. Furthermore, the immunodevice was used to screen for Potato virus X in 10 samples from potato tubers and leaves. The paper-disc-interfaced immunodevice was also evaluated by detecting other biomarkers, such as potato aucuba mosaic virus in plant diseases and C-reactive protein in human ones. This immunodevice may allow the on-site monitoring of diverse biomarkers by simplifying the current point of care diagnostic tools.

Reliability of Weight-Bearing Ultrasonography for Diagnosis of Lisfranc Joint Injuries: A Cadaveric Study

Category: Trauma Introduction/Purpose: Ultrasonography is increasingly used to evaluate the stability of various joint in the field of foot and ankle. This study aims to assess and compare the Lisfranc joint stability under stimulated weight bearing condition among progressive stages of Lisfranc ligamentous complex injuries. All the measurements were assessed using ultrasonography, fluoroscopy, and direct open measurement. We hypothesize that there is a correlation between ultrasonographic and fluoroscopic measurements, or between ultrasonographic and direct open measurements when performing stress evaluation of the Lisfranc instability. Methods: Twelve fresh below-knee amputated cadaveric specimens underwent ultrasound, fluoroscopic, and direct gross evaluation of the Lisfranc stability under simulated weight bearing condition. The assessment was first performed with all Lisfranc ligamentous complex intact and thereafter with sequential transection of the dorsal Lisfranc ligament, interosseous Lisfranc ligament, and plantar Lisfranc ligament. The simulated weight bearing condition was performed with 600N of force. The distances evaluated during the experimental set up were C1-M1, C2-M2, C1-M2, C1-C2 and M1-M2. All measurements were recorded in millimeter (mm). The ultrasound machine (GE, LOGIQ E-9 ultrasound machine, Chicago, Illinois, USA) was used to assess the Lisfranc stability by measuring the distances between the hyperechoic bone contours or the pre-defined landmark of the metatarsal and cuneiform bones. Correlations between 1) Ultrasound and fluoroscopic measurements, 2) Ultrasound and direct open measurements, and 3) Fluoroscopic and direct open measurements were evaluated using Pearson correlations coefficient. Results: Strong positive correlations between ultrasonographic and direct open measurements were found when assessed for C1-M1, C1-M2, C1-C2, M1-M2 (Pearson's correlation ranged from 0.73-0.97, p-values <0.001). Strong positive correlations between ultrasonographic and fluoroscopic measurements were found when assessed for C1-M1 from dorsal, medial, and plantar aspect (Pearson's correlation ranged from 0.75-0.90, p-values <0.001). Strong positive correlations between fluoroscopic and direct open measurements were found when assessed for C1-M1, C1-M2, C2-M2, M1-M2 (Pearson's correlation ranged from 0.77-0.93, p-values <0.001). Inter-rater and intra-rater reliability for the ultrasonographic measurements between the MSK radiologist (TK) and the foot and ankle surgeon (JS) were substantial (ICC ranged from 0.81-0.98). Conclusion: Ultrasonography is a reliable and reproducible tool for diagnosing Lisfranc ligamentous complex injuries. Both ultrasonographic and fluoroscopic measurements appear to strongly correlate with values measured by direct open measurement. Based on these data, ultrasonography is likely to become a valuable point of care diagnostic tool due to its ability to readily and dynamically evaluate suspected Lisfranc ligamentous complex injuries, without radiation exposure.

Intracranial pressure and veins.

This review summarizes the relationship between intracranial pressure and the venous system as far as ultrasound methods are concerned. Although the pressure in cortical veins is tightly and linearly coupled to the intracranial pressure, venous flow velocities in the basal veins are not. However, venous flow velocities reflect better the local cerebral blood flow than measurements in arteries. Therefore, they correlate better with clinical outcome in head trauma and subarachnoid hemorrhage. Transorbital sonography with measurement of the optic nerve sheath diameter and optic disk elevation has evolved to a valuable point of care diagnostic tool in idiopathic intracranial hypertension. In cerebral vein and sinus thrombosis ultrasound is at best a supplementary diagnostic tool, which may have a value in patient follow-up.

Measuring the sero-prevalence of Leishmania donovani induced cutaneous leishmaniasis: A method comparison study

An in-house enzyme-linked immunosorbent assay (ELISA) based on crude antigen of Leishmania reported a high sero-prevalence (82.0%) in Leishmania donovani induced cutaneous leishmaniasis (CL) in Sri Lanka. ELISA was further compared with established serological tools to identify a suitable point of care diagnostic tool. Sero-prevalence of 100 CL samples were analyzed using in-house ELISA, Indian dipstick test and rK39 strip test. Results obtained were further compared with direct agglutination test (DAT) for 40 CL. Test performance was evaluated using Kappa index value. Clinico-epidemiological characteristics of patients were analyzed using SPSSv25.0. Cost analysis of tests was carried out. ELISA showed a high sero-positivity of 81.0% (n = 81/100) while DAT (57.5%,n = 23/40), Indian dipstick test (22.0%,n = 22/100) and rK39 test (15.0%,n = 15/100) showed a comparatively less sero-positivity. According to Kappa index values, there were no perfect agreement between tests. Among ELISA positive patients (n = 81/100), DAT, Indian dipstick test and rK39 demonstrated sero-positivity rates of 61.3% (n = 19/31), 25.9% (n = 21/81) and 16.0% (n = 13/81) respectively. Among ELISA negative patients (n = 19/100), three assays demonstrated sero-positivity rates of 44.4% (n = 4/9), 5.3% (n = 1/9) and 10.5% (n = 2/19) respectively. DAT can be used as an alternative test when ELISA is not available or negative. Clinico-epidemiological profiles of patients that showed sero-positivity by each assay were different. Cost per patient was approximately 5.5 USD for DAT and 3.0 USD for each other tests. Established serological tests demonstrated different and relatively lower detection rates. Species, strain and antigen heterogeneity, inconsistency in amount of used antigens, sera, antibody expression patterns and testing methodologies could be responsible. This study highlighted the importance of designing an in-house serological assay based on local parasite.

Clinical testing on SARS-CoV-2 swab samples using reverse-transcription loop-mediated isothermal amplification (RT-LAMP)

BackgroundHigh cost of commercial RNA extraction kits limits the testing efficiency of SARS-CoV-2. Here, we developed a simple nucleic acid extraction method for the detection of SARS-CoV-2 directly from nasopharyngeal swab samples.MethodsA pH sensitive dye was used as the end point detection method. The obvious colour changes between positive and negative reactions eliminates the need of other equipment.ResultsClinical testing using 260 samples showed 92.7% sensitivity (95% CI 87.3–96.3%) and 93.6% specificity (95% CI 87.3–97.4%) of RT-LAMP.ConclusionsThe simple RNA extraction method minimizes the need for any extensive laboratory set-up. We suggest combining this simple nucleic acid extraction method and RT-LAMP technology as the point-of care diagnostic tool.

Ultrasensitive electrochemical genosensors for species-specific diagnosis of malaria

The absence of reliable species-specific diagnostic tools for malaria at point-of-care (POC) remains a major setback towards effective disease management. This is partly due to the limited sensitivity and specificity of the current malaria POC diagnostic kits especially in cases of low-density parasitaemia and mixed species infections. In this study, we describe the first label-free DNA-based genosensors based on electrochemical impedance spectroscopy (EIS) for species-specific detection of P. falciparum, P. malariae and P. ovale. The limits of detection (LOD) for the three species-specific genosensors were down in attomolar concentrations ranging from 18.7 aM to 43.6 aM, which is below the detection limits of previously reported malaria genosensors. More importantly, the diagnostic performance of the three genosensors were compared to quantitative real-time polymerase chain reaction (qPCR) assays using purified genomic DNA and the paired whole blood lysates from clinical samples. Remarkably, all the qPCR-positive purified genomic DNA samples were correctly identified by the genosensors indicating 100% sensitivity for each of the three malaria species. The specificities of the three genosensors ranged from 66.7% to 100.0% with a Therapeutic Turnaround Time (TTAT) within 30 min, which is comparable to the TTAT of current POC diagnostic tools for malaria. This work represents a significant step towards the development of accurate and rapid species-specific nucleic acid-based toolkits for the diagnosis of malaria at the POC.

Field evaluation of a novel, rapid diagnostic assay, and molecular epidemiology of enterotoxigenic E. coli among Zambian children presenting with diarrhea.

BackgroundEnterotoxigenic Escherichia coli (ETEC) is one of the top aetiologic agents of diarrhea in children under the age of 5 in low-middle income countries (LMICs). The lack of point of care diagnostic tools for routine ETEC diagnosis results in limited data regarding the actual burden and epidemiology in the endemic areas. We evaluated performance of the novel Rapid LAMP based Diagnostic Test (RLDT) for detection of ETEC in stool as a point of care diagnostic assay in a resource-limited setting.MethodsWe conducted a cross-sectional study of 324 randomly selected stool samples from children under 5 presenting with moderate to severe diarrhea (MSD). The samples were collected between November 2012 to September 2013 at selected health facilities in Zambia. The RLDT was evaluated by targeting three ETEC toxin genes [heat labile toxin (LT) and heat stable toxins (STh and STp)]. Quantitative PCR was used as the “gold standard” to evaluate the diagnostic sensitivity and specificity of RLDT for detection of ETEC. We additionally described the prevalence and seasonality of ETEC.ResultsThe study included 324 participants, 50.6% of which were female. The overall prevalence of ETEC was 19.8% by qPCR and 19.4% by RLDT. The children between 12 to 59 months had the highest prevalence of 22%. The study determined ETEC toxin distribution was LT 28/321(9%), ST 18/321(6%) and LT/ST 16/321(5%). The sensitivity and specificity of the RLDT compared to qPCR using a Ct 35 as the cut-off, were 90.7% and 97.5% for LT, 85.2% and 99.3% for STh and 100% and 99.7% for STp, respectively.ConclusionThe results of this study suggest that RLDT is sufficiently sensitive and specific and easy to implement in the endemic countries. Being rapid and simple, the RLDT also presents as an attractive tool for point-of-care testing at the health facilities and laboratories in the resource-limited settings.

FTIR spectroscopy as a point of care diagnostic tool for diabetes and periodontitis: A saliva analysis approach

Diabetes mellitus (DM) is a general term for heterogeneous metabolic disorders whose main characteristic is chronic hyperglycemia. Considering that conventional diagnostic methods are currently unable of early DM detection and the number of diabetic patients has been increasing worldwide, there is a clinical need for novel diagnostic approaches. Fourier-transform infrared (FT-IR) spectroscopy technique an alternative non-invasive diagnostic method for real-time evaluation of biofluids such as saliva. This study aims evaluate the feasibility of diagnosing diabetes and periodontitis through saliva samples based on their FT-IR spectra. Our first collection and spectral analysis of samples was a pilot study and comprised a total of 23 patients, 2 healthy, 9 with diabetes and 12 with diabetes and periodontitis. By using weighted KNN as classifier, we have obtained an area under the Receiver Operating Characteristic curve of 0.92 and 0.95 when considering the diabetes or diabetes + periodontitis groups as positive groups, respectively.

Evaluation of the HemoCue® WBC System as a Point of Care Diagnostic Tool for White Blood Cell Quantification in Pinnipeds

Point of care (POC) hematology testing can be valuable in triage and field settings. We assessed the accuracy between the HemoCue® WBC system and two comparative analyzers, as well as the precision of the HemoCue® WBC system in five different pinniped species: Zalophus californianus, Arctocephalus townsendi, Callorhinus urcinus, Phoca vitulina, and Mirounga angustirostris for white blood cell (WBC) quantification. In Zalophus (n = 164; 106 from U.S. Navy Marine Mammal Program (Navy); 58 from The Marine Mammal Center (TMMC)), the HemoCue® was compared to two hematology analyzers, Sysmex Xe-5000 and Vet ABC Plus. In Phoca (n = 50; TMMC), Callorhinus (n = 29; TMMC), Arctocephalus (n = 17; TMMC), and Mirounga (n = 67; TMMC), the HemoCue® was compared to Vet ABC Plus only. Bland–Altman and Passing–Bablok agreement of HemoCue® with Sysmex Xe-5000 and Vet ABC Plus analyzers were good for Zalophus, Arctocephalus, Phoca, and Mirounga but marginal with Callorhinus; bias = 0.56 × 109/L (Zalophus; Navy), −2.13 × 109/L (Zalophus; TMMC), −1.59 × 109/L (Arctocephalus), −2.48 × 109/L 0.917 (Phoca), −0.01 × 109/L (Mirounga), and −6.05 × 109/L (Callorhinus). The coefficient of variation from triplicate runs of samples were within acceptable limits for all species (2.50% ± 1.63 (Zalophus; TMMC), 3.09% ± 2.14 (Arctocephalus), 2.47% ± 1.35 (Callorhinus), 2.88% ± 1.75 (Phoca), and 3.44% ± 2.53 (Mirounga)), respectively. The presence of nucleated red blood cells (nRBC; 1–37 nRBC/100 WBC) did not significantly interfere with WBC counts in Zalophus, Callorhinus, and Phoca at the population level, but their presence should be evaluated at the individual level. The HemoCue® provides an accurate method for WBC quantification with WBC counts up to 30 × 109/L (upper limit of linearity of the analyzer) in Zalophus, Arctocephalus, Phoca, and Mirounga, but is less accurate in Callorhinus, and showed good precision in all species.

Plasmonic nanosensors for point-of-care biomarker detection.

Advancement of materials along with their fascinating properties play increasingly important role in facilitating the rapid progress in medicine. An excellent example is the recent development of biosensors based on nanomaterials that induce surface plasmon effect for screening biomarkers of various diseases ranging from cancer to Covid-19. The recent global pandemic re-confirmed the trend of real-time diagnosis in public health to be in point-of-care (POC) settings that can screen interested biomarkers at home, or literally anywhere else, at any time. Plasmonic biosensors, thanks to its versatile designs and extraordinary sensitivities, can be scaled into small and portable devices for POC diagnostic tools. In the meantime, efforts are being made to speed up, simplify and lower the cost of the signal readout process including converting the conventional heavy laboratory instruments into lightweight handheld devices. This article reviews the recent progress on the design of plasmonic nanomaterial-based biosensors for biomarker detection with a perspective of POC applications. After briefly introducing the plasmonic detection working mechanisms and devices, the selected highlights in the field focusing on the technology's design including nanomaterials development, structure assembly, and target applications are presented and analyzed. In parallel, discussions on the sensor's current or potential applicability in POC diagnosis are provided. Finally, challenges and opportunities in plasmonic biosensor for biomarker detection, such as the current Covid-19 pandemic and its testing using plasmonic biosensor and incorporation of machine learning algorithms are discussed.

Use of portable ultrasonography for the diagnosis of lateral ankle instability.

Portable ultrasonography is increasingly used to evaluate ankle stability at the point of care. This study aims to determine the correlation of portable-ultrasonographic and fluoroscopic measurements of ankle laxity in a cadaveric ligament transection model of ankle ligament injury. We hypothesize that there is an association between portable-ultrasonographic and fluoroscopic measurements when performing stress evaluation of lateral ankle instability. Eight fresh-frozen below-knee amputated cadaveric specimens with intact proximal fibula underwent ultrasound and fluoroscopic evaluation of the ankle during anterior drawer and talar tilt testing. The assessment was first performed with all lateral ankle ligaments intact and thereafter with sequential transection of the anterior talofibular ligament, calcaneofibular ligament, and posterior talofibular ligament. The anterior drawer test was performed with both 50N and 80N of force, and talar tilt test was performed with 1.7 Nm of torque. Correlations between (1) portable-ultrasonographic and fluoroscopic measurements and (2) sequential transection of lateral ankle ligaments were evaluated using Spearman's rank correlations. The same statistical test was used to investigate the correlation between the ultrasonographic and the fluoroscopic measurements. The inter- and intra-observer agreement was assessed using the intraclass correlation coefficient through a two-way mixed-effects model with absolute agreement. Portable-ultrasonographic and fluoroscopic measurements increased as additional ligaments of the lateral ankle were transected (Spearman's rank correlation ranged from 0.74 to 0.81, 0.74 to 0.81, p-values < 0.001). Strong positive correlations between ultrasonographic and fluoroscopic measurements were found during the lateral ankle stability evaluation using anterior drawer and talar tilt testing (Spearman's rank correlation ranged from 0.81 to 0.85, 0.81 to 0.85, p-values < 0.001). Inter-rater (0.99, 95% CI: 0.98-0.99) and intra-rater reliability (0.97, 95% CI: 0.95-0.99) for the ultrasonographic measurements were substantial. In conclusion, there was a strong correlation found between ultrasonographic and fluoroscopic values measured during simulated anterior drawer and talar tilt test in a cadaveric ligament transection model. In this model, the portable-ultrasonographic measurement was found to be reliable for repeated measurements of the talar translation and the lateral clear space distance. Based on these data, ultrasonography is likely to become a valuable point of care diagnostic tool due to its ability to readily and dynamically evaluate suspected lateral ankle instability. Clinical Significance: The use of dynamic stress ultrasound to assess the anterior translation of the talus and the lateral clear space distance appears to be a reliable and repeatable technique to evaluate lateral ankle stability with a radiation-free, noninvasive, and low-cost manner.

Synthetic Biology Construct of Ebola Virus in Bacteria Surrogate Is Stable and Safe for Rapid Detection Studies in a BSL-2 Laboratory Setting

Rapid detection of virulent pathogens during an outbreak is critical for public health advisories and control of the disease in a population. While many molecular techniques for point of care and clinical diagnosis abound, the US experience with the COVID-19 testing in the early stages of the pandemic underscores the critical importance of determining the appropriate target gene(s) with in-built controls that reliably detect pathogens with high sensitivity and specificity. Assays and research for diagnostics and therapy could be slowed during an epidemic because access to the required BSL-3 and BSL-4 laboratories are limited. So, during the 2014 West Africa Ebola outbreak, we tested the hypothesis that using synthetic cDNA of Ebolavirus in a bacteria surrogate (fit for all lab settings), would remain unmutated and safe after several generations, serving as an effective positive control in research settings, self test and point-of-care detection platforms. Primers were designed for the detection and quantification of the nucleoprotein (NP) gene of the 2014 Makona Ebola strain (KR781608.1, 733 - 1332 bp). To test the stability of artificially inserted translation arrest in the Orf of the model gene, it was edited to include three STOP codons in the RNA transcript using SNAP GENE. The segment was then spliced into a high copy number plasmid, cloned into One ShotTM TOP10 Escherichia coli (Invitrogen), and tested for stability and safety by periodic subculture, extraction and sequencing. Unlike COVID-19, rapid detection of blood-borne etiologies like Ebola requires optimized protocols for blood matrix. Using real-time PCR and newly designed primer pairs, the EBOV surrogate was detected and enumerated in human blood and regular broth and buffers. Based on aligned sequence analysis, the EBOV synthetic NP gene was stable (>99.9999% similarity coefficient) for at least 3 months. Detection sensitivity in broth and blood was at least 100 cells/ml or about 5.8 × 103 to 7.3 × 103 virion equivalents per ml. While the developments of transcription-and-replication-competent virus like particles (trVLP) have made it possible to study the infection and replication cycles of virulent pathogens in BSL-2 laboratories, the simplicity of our model and the reproducibility of detection and enumeration show the utility of synthetic bio-components as positive controls for point of care diagnostic tools. The inserted stop codons remained intact after many generations, suggesting that expressed virulent proteins can be easily silenced in synthetic biology models for research in BSL-1 and 2 and a wide range of pathogens. Synthetic bio-components can thereby aid further research by reducing costs and improving safety for workers and stakeholders.

Novel Biorecognition Elements against Pathogens in the Design of State-of-the-Art Diagnostics.

Infectious agents, especially bacteria and viruses, account for a vast number of hospitalisations and mortality worldwide. Providing effective and timely diagnostics for the multiplicity of infectious diseases is challenging. Conventional diagnostic solutions, although technologically advanced, are highly complex and often inaccessible in resource-limited settings. An alternative strategy involves convenient rapid diagnostics which can be easily administered at the point-of-care (POC) and at low cost without sacrificing reliability. Biosensors and other rapid POC diagnostic tools which require biorecognition elements to precisely identify the causative pathogen are being developed. The effectiveness of these devices is highly dependent on their biorecognition capabilities. Naturally occurring biorecognition elements include antibodies, bacteriophages and enzymes. Recently, modified molecules such as DNAzymes, peptide nucleic acids and molecules which suffer a selective screening like aptamers and peptides are gaining interest for their biorecognition capabilities and other advantages over purely natural ones, such as robustness and lower production costs. Antimicrobials with a broad-spectrum activity against pathogens, such as antibiotics, are also used in dual diagnostic and therapeutic strategies. Other successful pathogen identification strategies use chemical ligands, molecularly imprinted polymers and Clustered Regularly Interspaced Short Palindromic Repeats-associated nuclease. Herein, the latest developments regarding biorecognition elements and strategies to use them in the design of new biosensors for pathogens detection are reviewed.

Rapid and sensitive fluorescence and smartphone dual-mode detection of dopamine based on nitrogen-boron co-doped carbon quantum dots

A dual-mode fluorescence and colorimetric biosensor based on nitrogen-boron co-doped carbon quantum dot (N-B CQDs) for rapid and sensitive detection of dopamine (DA) was developed. The quantum dot luminescent materials, N-B CQDs, were prepared by a one-step microwave-assisted method. The N-B CQDs were characterized using SEM, HR-TEM, XRD, FT-IR, Raman, fluorescence, and UV-Vis techniques. The dual-mode assays of fluorescence and colorimetric methods were used for detection of DA. The high fluorescent N-B CQDs mediated turn-off assay for the facile room temperature detection of dopamine via inner filter effect (IFE) and Forster resonance energy transfer (FRET) processes atbasic pH. The colorimetric detection of DA was also developed via in-house android application using a smartphone and N-B CQD solution-based nanosensor. The smartphone-based colorimetric biosensors generated morereliable information for quantitative analysis of color changes than the naked eye. Furthermore, a smartphone application with N-B CQD solution-based nanosensor was integrated to monitor the color changes through the DA addition. Wide linear ranges were achieved for DA in the ranges 0.25-50μM and 5-500μM with fluorescence and smartphone-based method, respectively. The satisfactory results of the dual-mode detection of DA, not only in aqueous solution, but also in human urine and serum biological sample demonstrated its potential application in biosensing, asa point of care diagnostic tool. Graphical abstract.

Microfluidics as an Emerging Platform for Tackling Antimicrobial Resistance (AMR): A Review

Background: Antimicrobial resistance (AMR) occurs when microbes become resistant to antibiotics causing complications and limited treatment options. AMR is more significant where antibiotics use is excessive or abusive and the strains of bacteria become resistant to antibiotic treatments. Current technologies for bacteria and its resistant strains identification and antimicrobial susceptibility testing (AST) are mostly central-lab based in hospitals, which normally take days to weeks to get results. These tools and procedures are expensive, laborious and skills based. There is an ever-increasing demand for developing point-of-care (POC) diagnostics tools for rapid and near patient AMR testing. Microfluidics, an important and fundamental technique to develop POC devices, has been utilized to tackle AMR in healthcare. This review mainly focuses on the current development in the field of microfluidics for rapid AMR testing. Method: Due to the limitations of conventional AMR techniques, microfluidic-based platforms have been developed for better understandings of bacterial resistance, smart AST and minimum inhibitory concentration (MIC) testing tools and development of new drugs. This review aims to summarize the recent development of AST and MIC testing tools in different formats of microfluidics technology. Results: Various microfluidics devices have been developed to combat AMR. Miniaturization and integration of different tools has been attempted to produce handheld or standalone devices for rapid AMR testing using different formats of microfluidics technology such as active microfluidics, droplet microfluidics, paper microfluidics and capillary-driven microfluidics. Conclusion: Current conventional AMR detection technologies provide time-consuming, costly, labor-intensive and central lab-based solutions, limiting their applications. Microfluidics has been developed for decades and the technology has emerged as a powerful tool for POC diagnostics of antimicrobial resistance in healthcare providing, simple, robust, cost-effective and portable diagnostics. The success has been reported in research articles; however, the potential of microfluidics technology in tackling AMR has not been fully achieved in clinical settings.

The role of vascular ultrasound in managing giant cell arteritis in ophthalmology

Giant cell arteritis is the most common systemic vasculitis in the elderly and is a potentially life-threatening ophthalmic emergency that can result in irreversible blindness. Blindness is most commonly associated with acute onset, irreversible arteritic ischemic optic neuropathy. Without treatment, second eye involvement may occur, resulting in bilateral blindness. Patients with established visual loss are treated with high-dose steroids and generally undergo a temporal artery biopsy to confirm their diagnosis. A significant number of patients are, however, referred for urgent ophthalmology assessment from concerns about “incipient” arteritic ischemic optic neuropathy. Before visual loss, patients may experience a range of ocular symptoms related to ischemia. This generally leads to treatment with high-dose systemic steroid and an urgent request for a temporal artery biopsy. Temporal artery biopsy is considered as the standard investigation for confirmatory diagnosis. It is generally arranged as soon as possible, although it is often not carried out for several days, and there may also be delays in histopathological reporting. It is often perceived that the patient is “safe” while on corticosteroids, in that they are being treated to avoid visual loss. What is not acknowledged, however, is that, if patients do not have giant cell arteritis and are being treated “just in case,” they will often require a tapering of oral steroids over several weeks, exposing them to unnecessary and significant side effects. In the rheumatology setting, vascular ultrasound has emerged as a safe and reliable alternative to temporal artery biopsy as a point of care diagnostic tool in the management of giant cell arteritis. Given an experienced sonographer and optimal equipment, a rapid diagnosis can be established in a fast-track clinic setting, taking into consideration clinical assessment, scoring, and ultrasound findings. A huge advantage of ultrasound is that it provides immediate information that can be used to inform treatment decisions. We explore the evidence that supports the incorporation of vascular ultrasound into the ophthalmology repertoire to make a more efficient diagnosis that is cost-effective and associated with better patient outcomes, including a potential reduction in loss of sight and avoidance of unnecessary long-term steroid treatment by early exclusion of mimics.

Advances in Directly Amplifying Nucleic Acids from Complex Samples.

Advances in nucleic acid amplification technologies have revolutionized diagnostics for systemic, inherited, and infectious diseases. Current assays and platforms, however, often require lengthy experimental procedures and multiple instruments to remove contaminants and inhibitors from clinically-relevant, complex samples. This requirement of sample preparation has been a bottleneck for using nucleic acid amplification tests (NAATs) at the point of care (POC), though advances in “lab-on-chip” platforms that integrate sample preparation and NAATs have made great strides in this space. Alternatively, direct NAATs—techniques that minimize or even bypass sample preparation—present promising strategies for developing POC diagnostic tools for analyzing real-world samples. In this review, we discuss the current status of direct NAATs. Specifically, we surveyed potential testing systems published from 1989 to 2017, and analyzed their performances in terms of robustness, sensitivity, clinical relevance, and suitability for POC diagnostics. We introduce bubble plots to facilitate our analysis, as bubble plots enable effective visualization of the performances of these direct NAATs. Through our review, we hope to initiate an in-depth examination of direct NAATs and their potential for realizing POC diagnostics, and ultimately transformative technologies that can further enhance healthcare.