Search Results (24)

Accurate and rapid detection of Plasmodium falciparum parasites in blood smears is critical for the timely diagnosis and treatment of malaria, particularly in resource-constrained field settings. This paper presents a proof-of-concept solution demonstrating the feasibility of the Google Coral Edge Tensor Processing Unit (TPU) for real-time screening of thin blood smears for P. falciparum infection. We develop and deploy a lightweight deep learning model optimized for edge inference using transfer learning and training data supplied by the NIH. This model is capable of detecting individual parasitized red blood cells (RBCs) with high sensitivity and specificity. In a final deployment, the system will integrate a portable digital microscope and low-power color display with the Coral TPU to perform on-site image capture and classification without reliance on cloud connectivity. We detail the model training process using a curated dataset of annotated smear images, potential future hardware integration for field deployment, and performance benchmarks. Initial tests show that the Coral TPU-based solution achieves an accuracy of 92% in detecting P. falciparum parasites in thin-smear microscopy images, with processing times under 50 ms per identified RBC. This work illustrates the potential of edge AI devices to transform malaria diagnostics in low-resource settings through efficient, affordable, and scalable screening tools. Full article

Simultaneous molecular recognition and quantification of at least four biomarkers in biological samples may contribute to early and fast diagnosis of illnesses such as cancer. The electrodes able to reliably perform on-site these tests are the stochastic sensors. Therefore, three novel 3D stochastic sensors employing carbon-based powders (graphite, graphene, nanographene) treated with N-(2-mercapto-1H-benzo[d]imidazole-5-yl) oleamide solution were used for screening tests of whole blood, gastric tumoral tissue, urine, and saliva for molecular recognition and quantification of CA12-5, CA72-4, HER1, and AFP. The best performance was achieved for the sensor based on graphene, when the highest sensitivities were recorded, on wide working concentration ranges of: 8.37 × 10⁻¹⁴–8.37 U mL⁻¹ for CA12-5, 4.00 × 10⁻¹¹–4.00 × 10⁻³ U mL⁻¹ for CA72-4, 3.90 × 10⁻¹⁶–3.90 × 10⁻⁶ g mL⁻¹ for HER1, and 3.00 × 10⁻²⁰–3.00 × 10⁻⁶ g mL⁻¹ for AFP. The wide linear concentration ranges cover levels of biomarkers found in gastric cancer patients from early to late stages. The recovery values were higher than 98.00 with %, RSD lower than 1.00%. Full article

Pigs are susceptible to the deadly infectious disease known as African swine fever (ASF), which is brought on by the African swine fever virus (ASFV). As such, prompt and precise disease detection is essential. Deletion of the virulence-related genes MGF-505/360 and EP402R generated from the virulent genotype II virus significantly reduces its virulence, and animal tests using one of the recombinant viruses show great lethality and transmissibility in pigs. The isothermal technique known as recombinase polymerase amplification (RPA) is perfect for rapid in-field detection. To accurately identify ASFV MGF-505R gene-deleted mutants and assess the complex infection situation of ASF, RPA assays in conjunction with real-time fluorescent detection (real-time RPA assay) and lateral flow dipstick (RPA-LFD assay) were created. These innovative methods allow for the direct detection of ASFV from pigs, offering in-field pathogen detection, timely disease management, and satisfying animal quarantine requirements. The specific primers and probes were designed against conserved regions of ASFV B646L and MGF-505R genes. Using recombinant plasmid DNA containing ASFV MGF-505R gene-deleted mutants as a template, the sensitivity of both ASF real-time RPA and ASF RPA-LFD assays were demonstrated to be 10 copies per reaction within 20 min at 37 °C. Neither assay had cross-reactions with CSFV, PRRSV, PPV, PRV, ot PCV2, common viruses seen in pigs, indicating that these methods were highly specific for ASFV. The evaluation of the performance of ASFV real-time RPA and ASFV RPA-LFD assays with clinical samples (n = 453) demonstrated their ability to specifically detect ASFV or MGF-505R gene-deleted mutants in samples of pig feces, ham, fresh pork, and blood. Both assays exhibited the same diagnostic rate as the WOAH-recommended real-time fluorescence PCR, highlighting their reliability and validity. These assays offer a simple, cost-effective, rapid, and sensitive method for on-site identification of ASFV MGF-505R gene-deleted mutants. As a promising alternative to real-time PCR, they have the potential to significantly enhance the prevention and control of ASF in field settings. Full article

Voluntary HIV testing and counselling (VCT) in the workplace could reach population groups who may be at risk for HIV but may not readily seek out testing from other services. We conducted a scoping review to understand (a) the nature of evidence related to initiatives and interventions for vocationally active adults on VCT in occupational settings, and (b) any facilitators and barriers to the delivery of and/or engagement with VCT initiatives/interventions in the workplace. JBI scoping review methodology was followed. The protocol was pre-registered. Included studies focused on vocationally active adults (population), VCT interventions or initiatives (concept), and workplaces in any sector or country (context). The review included studies published after 2000, in English, and of any research design. Studies relating to mandatory workplace HIV screening were excluded. MEDLINE, CINAHL, Scopus, PsycINFO, and the Cochrane Central Register of Control Trials were searched. Sources of grey literature included Google Scholar and governmental and organisational websites. One reviewer screened titles and abstracts; a second reviewer independently screened 10%. Data extraction utilised a modified JBI data extraction tool. We identified 17 studies reporting on 12 workplace VCT interventions (20,985 participants, 15–70 years). Studies were conducted in eight countries between 2001 and 2022. Interventions were delivered in organisations of different types, sizes and sectors. Testing included rapid blood tests and oral fluid self-tests. Where reported, the average on-site HIV testing uptake rate was 63%, and the average linkage to care rate was 86.85%. Views of workers, employers and service providers were largely positive. Barriers included being male, masculinity-driven workplace culture, HIV-related stigma, poor knowledge, low risk perceptions, lack of time and low support. Facilitators included on-site testing for convenience and accessibility, rapid and free tests, organisational, managerial and peer support, and embedding HIV tests within general health checks. Evaluation methods varied, although randomised trial designs were uncommon. Despite the limited number of studies, the workplace appears to be a viable route to the delivery of community-based VCT, albeit barriers should be addressed. Reporting quality of interventions and associated evaluations is variable and could be improved with the use of appropriate checklists. Full article

The calcium requirements of dairy cows increase dramatically soon after calving, and many cows have subclinical hypocalcemia, adversely affecting health and performance. Traditional laboratory tests for calcium are complex and not easily adapted to rapid point-of-care applications. The objectives were to evaluate a portable iCa testing device, Horiba LAQUAtwin Ca-11C, for measuring ionized calcium (iCa) in the whole blood of dairy cows and to investigate the iCa-to-total-calcium (tCa) ratio in blood collected from dairy cows within 9 days after calving. This study was conducted on two large dairies in northern China. First, blood samples were collected from 246 cows within 3 days after calving, and whole-blood iCa concentrations were measured with a Horiba LAQUAtwin Ca-11C and Abbott i-STAT 1. In addition, 885 blood samples were collected from 102 dairy cows between calving and 9 days postpartum, with iCa concentrations measured using a Horiba LAQUAtwin Ca-11C and tCa concentrations determined by a fully automated biochemical analyzer. The Horiba LAQUAtwin Ca-11C and Abbott i-STAT 1 had significant consistency in the measurement of ionized calcium concentration in whole blood (the frequency of differences between the measured values was within ±20% of the average, reaching 95.53%), and the measurement results of the two instruments were correlated (Deming regression analysis R² = 0.87). This implied the potential application of the Horiba LAQUAtwin Ca-11C as a simplified device for measuring iCa in dairy farms for on-site testing. Within the first 9 days after calving, the ratio of ionized calcium to total calcium fluctuated but overall remained within the range of 44.2 to 47.22%. Furthermore, significant variations in the ratio among individual cows indicated that individual differences and physiological states of cows affected calcium metabolism. This study identified the potential to use the Horiba LAQUAtwin Ca-11C for point-of-care testing on dairy farms. Nevertheless, the impacts of factors such as health status and individual cow differences on the ratio of ionized calcium to total calcium still require further investigation. Full article

In order to develop a yak/chauri-specific welfare assessment protocol, we sent a set of 31 potential welfare measures to 120 Nepalese experts and asked them to identify the measures that they thought would be useful and propose additional useful measures. Eighty-three experts responded, with 13 measures being identified by >50% of respondents as likely to be useful. These thirteen measures plus one new measure (hematology) were included in an assessment protocol that was tested in the second phase of this study in five chauri herds in two districts in northern Nepal. Animal-based evaluations along with sampling for mastitis, intestinal parasites, and hematology were undertaken during or just after morning milking. Resource- and record-based measures were assessed through structured interviews, with verifications on-site where possible. No chauris exhibited poor body conditions, skin injuries, significant locomotion issues, or significant subclinical mastitis. Fecal testing suggested a high prevalence of intestinal parasites at the herd level, while blood testing suggested no evidence of hematological abnormalities. However, for both results, we need more data to use these effectively as measures of welfare. The resource-based assessment revealed significant challenges across all resources, and veterinary services were reported as being inadequate. A high estimated annual mortality rate (10–21%) needs further investigation. This protocol provided a useful start towards developing a welfare assessment protocol for yak/chauri and identified issues that need addressing to optimize chauri welfare. Full article

Malaria is listed as one of the three most hazardous infectious diseases worldwide. Travelers and migrants passing through exit and entry ports are important sources of malaria pandemics globally. Developing accurate and rapid detection technology for malaria is important. Here, a novel hairpin-mediated amplification (HMA) technique was proposed for the detection of four Plasmodium species, including P. falciparum, P. vivax, P. malariae, and P. ovale. Based on the conserved nucleotide sequence of Plasmodium, specific primers and probes were designed for the HMA process, and the amplicon can be detected using lateral flow detection (LFD); the results can be read visually without specialized equipment. The specificity of HMA-LFD was evaluated using nucleic acids extracted from four different Plasmodium species and two virus species. The sensitivity of HMA-LFD was valued using 10× serial dilutions of plasmid containing the template sequence. Moreover, 78 blood samples were collected to compare HMA-LFD and qPCR. The HMA-LFD results were all positive for four different Plasmodium species and negative for the other two virus species. The sensitivity of HMA-LFD was tested to be near five copies/μL. The analysis of clinical samples indicated that the consistency of HMA-LFD and qPCR was approximately 96.15%. Based on these results, the HMA-LFD assay was demonstrated to be a rapid, sensitive, and specific technique for the detection of Plasmodium and has great advantages for on-site detection in low-resource areas and exit and entry ports. Full article

Background: Neonatal pain assessment (NPA) represents a huge global problem of essential importance, as a timely and accurate assessment of neonatal pain is indispensable for implementing pain management. Purpose: To investigate the consistency of pain scores derived through video-based NPA (VB-NPA) and on-site NPA (OS-NPA), providing the scientific foundation and feasibility of adopting VB-NPA results in a real-world scenario as the gold standard for neonatal pain in clinical studies and labels for artificial intelligence (AI)-based NPA (AI-NPA) applications. Setting: A total of 598 neonates were recruited from a pediatric hospital in China. Methods: This observational study recorded 598 neonates who underwent one of 10 painful procedures, including arterial blood sampling, heel blood sampling, fingertip blood sampling, intravenous injection, subcutaneous injection, peripheral intravenous cannulation, nasopharyngeal suctioning, retention enema, adhesive removal, and wound dressing. Two experienced nurses performed OS-NPA and VB-NPA at a 10-day interval through double-blind scoring using the Neonatal Infant Pain Scale to evaluate the pain level of the neonates. Intra-rater and inter-rater reliability were calculated and analyzed, and a paired samples t-test was used to explore the bias and consistency of the assessors’ pain scores derived through OS-NPA and VB-NPA. The impact of different label sources was evaluated using three state-of-the-art AI methods trained with labels given by OS-NPA and VB-NPA, respectively. Results: The intra-rater reliability of the same assessor was 0.976–0.983 across different times, as measured by the intraclass correlation coefficient. The inter-rater reliability was 0.983 for single measures and 0.992 for average measures. No significant differences were observed between the OS-NPA scores and the assessment of an independent VB-NPA assessor. The different label sources only caused a limited accuracy loss of 0.022–0.044 for the three AI methods. Conclusion: VB-NPA in a real-world scenario is an effective way to assess neonatal pain due to its high intra-rater and inter-rater reliability compared to OS-NPA and could be used for the labeling of large-scale NPA video databases for clinical studies and AI training. Full article

Human Mpox cases are increasingly reported in Africa, with the highest burden in the Democratic Republic of Congo (DRC). While case reporting on a clinical basis can overestimate infection rates, laboratory confirmation by PCR can underestimate them, especially on suboptimal samples like blood, commonly used in DRC. Here we used a Luminex-based assay to evaluate whether antibody testing can be complementary to confirm cases and to identify human transmission chains during outbreak investigations. We used left-over blood samples from 463 patients, collected during 174 outbreaks between 2013 and 2022, with corresponding Mpox and VZV PCR results. In total, 157 (33.9%) samples were orthopox-PCR positive and classified as Mpox+; 124 (26.8%) had antibodies to at least one of the three Mpox peptides. The proportion of antibody positive samples was significantly higher in Mpox positive samples (36.9%) versus negative (21.6%) (p < 0.001). By combining PCR and serology, 66 additional patients were identified, leading to an Mpox infection rate of 48.2% (223/463) versus 33.9% when only PCR positivity is considered. Mpox infections were as such identified in 14 additional health zones and 23 additional outbreaks (111/174 (63.8%) versus 88/174 (50.6%)). Our findings highlight the urgent need of rapid on-site diagnostics to circumvent Mpox spread. Full article

Point-of-care testing (POCT), also known as on-site or near-patient testing, has been exploding in the last 20 years. A favorable POCT device requires minimal sample handling (e.g., finger-prick samples, but plasma for analysis), minimal sample volume (e.g., one drop of blood), and very fast results. Shear horizontal surface acoustic wave (SH-SAW) biosensors have attracted a lot of attention as one of the effective solutions to complete whole blood measurements in less than 3 min, while providing a low-cost and small-sized device. This review provides an overview of the SH-SAW biosensor system that has been successfully commercialized for medical use. Three unique features of the system are a disposable test cartridge with an SH-SAW sensor chip, a mass-produced bio-coating, and a palm-sized reader. This paper first discusses the characteristics and performance of the SH-SAW sensor system. Subsequently, the method of cross-linking biomaterials and the analysis of SH-SAW real-time signals are investigated, and the detection range and detection limit are presented. Full article

Plasma separation from whole blood is oftent required as an essential first step when performing blood tests with a viral assay. However, developing a point-of-care plasma extraction device with a large output and high virus recovery remains a significant obstacle to the success of on-site viral load tests. Here, we report a portable, easy-to-use, cost-efficient, membrane-filtration-based plasma separation device that enables rapid large-volume plasma extraction from whole blood, designed for point-of-care virus assays. The plasma separation is realized by a low-fouling zwitterionic polyurethane-modified cellulose acetate (PCBU-CA) membrane. The zwitterionic coating on the cellulose acetate membrane can decrease surface protein adsorption by 60% and increase plasma permeation by 46% compared with a pristine membrane. The PCBU-CA membrane, with its ultralow-fouling properties, enables rapid plasma separation. The device can yield a total of 1.33 mL plasma from 10 mL whole blood in 10 min. The extracted plasma is cell-free and exhibits a low hemoglobin level. In addition, our device demonstrated a 57.8% T7 phage recovery in the separated plasma. The results of real-time polymerase chain reaction analysis confirmed that the nucleic acid amplification curve of the plasma extracted by our device is comparable to that obtained by centrifugation. With its high plasma yield and good phage recovery, our plasma separation device provides an excellent replacement for traditional plasma separation protocols for point-of-care virus assays and a broad spectrum of clinical tests. Full article

Methods based on nucleic acid detection are currently the most commonly used technique in COVID-19 diagnostics. Although generally considered adequate, these methods are characterised by quite a long time-to-result and the necessity to prepare the material taken from the examined person—RNA isolation. For this reason, new detection methods are being sought, especially those characterised by the high speed of the analysis process from the moment of sampling to the result. Currently, serological methods of detecting antibodies against the virus in the patient’s blood plasma have attracted much attention. Although they are less precise in determining the current infection, such methods shorten the analysis time to several minutes, making it possible to consider them a promising method for screening tests in people with suspected infection. The described study investigated the feasibility of a surface plasmon resonance (SPR)-based detection system for on-site COVID-19 diagnostics. A simple-to-use portable device was proposed for the fast detection of anti-SARS-CoV-2 antibodies in human plasma. SARS-CoV-2-positive and -negative patient blood plasma samples were investigated and compared with the ELISA test. The receptor-binding domain (RBD) of spike protein from SARS-CoV-2 was selected as a binding molecule for the study. Then, the process of antibody detection using this peptide was examined under laboratory conditions on a commercially available SPR device. The portable device was prepared and tested on plasma samples from humans. The results were compared with those obtained in the same patients using the reference diagnostic method. The detection system is effective in the detection of anti-SARS-CoV-2 with the detection limit of 40 ng/mL. It was shown that it is a portable device that can correctly examine human plasma samples within a 10 min timeframe. Full article

We propose a protocol suitable for point-of-care diagnosis of malaria utilizing a simple and purification-free DNA extraction method with the combination of loop-mediated isothermal amplification assay and lateral flow (LAMP-LF). The multiplex LAMP-LF platform developed here can simultaneously detect Plasmodium knowlesi, P. vivax, P. falciparum, and Plasmodium genus (for P. malariae and P. ovale). Through the capillary effect, the results can be observed by the red band signal on the test and control lines within 5 min. The developed multiplex LAMP-LF was tested with 86 clinical blood samples on-site at Hospital Kapit, Sarawak, Malaysia. By using microscopy as the reference method, the multiplex LAMP-LF showed 100% sensitivity (95% confidence interval (CI): 91.4 to 100.00%) and 97.8% specificity (95% CI: 88.2% to 99.9%). The high sensitivity and specificity of multiplex LAMP-LF make it ideal for use as a point-of-care diagnostic tool. The simple and purification-free DNA extraction protocol can be employed as an alternative DNA extraction method for malaria diagnosis in resource-limited settings. By combining the simple DNA extraction protocol and multiplex LAMP-LF approach, we aim to develop a simple-to-handle and easy-to-read molecular diagnostic tool for malaria in both laboratory and on-site settings. Full article

Rats impress by their high platelet count resulting in hypercoagulability, which protects the animals from severe bleeding. However, platelets also import numerous stiff junction points into the fibrous system of a clot, also enhancing the pre-stress of the fibrin fibers, which lowers their deformability. Clot deformation is clinically important since large strains are present in the arterial tree (caused by the propagation of pressure and pulse waves), and a clot is considered “safe” when it can deform over a long range of strain amplitudes. We tested clot formation and the behavior of fully formed blood clots of laboratory rats at large sinusoidal shear stress amplitudes by rheometry and compared outcomes to human reference data. We found that fiber density (by scanning electron microscopy) and clot stiffness (by rheometry) was pronounced compared to humans and differed with sexual dimorphism and with rat strain. Using our large amplitude oscillation (LAOS) protocol, we detected that rat clots yielded with a frustrated attempt to stiffen instead of showing the macroscopic stiffening response that is typical for human clots. We attribute this behavior to the appearance of multiple microfractures until, finally, a few leading fibers uptake the load. Rat clots also failed to align fibers in shear direction to initiate affine deformation. The rat clot phenotype differs substantially from the human one, which must be considered in research and toxicological testing. If microfractures in the fiber meshwork are concentrated in vivo, parts of a clot may break off and be washed away. However, homogenously distributed microfractures may open pores and allow the penetration of plasminogen activators. What occurs in the rat vasculature depends on the on-site clot composition. Full article

The development of precise and efficient diagnostic tools enables early treatment and proper isolation of infected individuals, hence limiting the spread of coronavirus disease 2019 (COVID-19). The standard diagnostic tests used by healthcare workers to diagnose severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection have some limitations, including longer detection time, the need for qualified individuals, and the use of sophisticated bench-top equipment, which limit their use for rapid SARS-CoV-2 assessment. Advances in sensor technology have renewed the interest in electrochemical biosensors miniaturization, which provide improved diagnostic qualities such as rapid response, simplicity of operation, portability, and readiness for on-site screening of infection. This review gives a condensed overview of the current electrochemical sensing platform strategies for SARS-CoV-2 detection in clinical samples. The fundamentals of fabricating electrochemical biosensors, such as the chosen electrode materials, electrochemical transducing techniques, and sensitive biorecognition molecules, are thoroughly discussed in this paper. Furthermore, we summarised electrochemical biosensors detection strategies and their analytical performance on diverse clinical samples, including saliva, blood, and nasopharyngeal swab. Finally, we address the employment of miniaturized electrochemical biosensors integrated with microfluidic technology in viral electrochemical biosensors, emphasizing its potential for on-site diagnostics applications. Full article