Search Results (18)

A series of incidents involving damage to vehicle speed sensor cables occurred in an urban area in Japan. At the request of the police, DNA analysis was conducted to identify the animal species responsible. Swab samples collected from the damaged sections of the cables were subjected to PCR testing using mtDNA fragments. Sequencing analysis with universal primers (SCPH02500, SCPL02981) detected DNA from the Japanese marten (Martes melampus). A comprehensive examination that included morphological analysis of the cable damage and consideration of the ecological characteristics of the Japanese martens suggested that the damage was likely caused by this species. DNA analysis using mtDNA markers is a valuable tool for species identification in wildlife forensic veterinary investigations and serves as important scientific evidence in criminal cases involving animals. The findings from this case may contribute to future investigations in forensic veterinary science and ecological research and may also inform measures to prevent human–wildlife conflicts involving animals. Full article

Mycoplasma pneumoniae (MP) is the main culprit of community-acquired pneumonia. Commonly used laboratory testing methods have many shortcomings. Serological diagnosis has low sensitivity, causing false negatives, while a quantitative real-time polymerase chain reaction (qPCR) requires large equipment and professional staff. To make up for these shortcomings, we proposed a label-free, low-cost, and small-sized ion-sensitive field-effect transistor (ISFET) array based on a low-buffered loop-mediated isothermal amplification (LAMP) assay. A complementary metal oxide semiconductor (CMOS)-based ISFET array with 512 × 512 sensors was used in this system, which responds specifically to H⁺ with a sensitivity of 365.7 mV/pH. For on-chip amplification, a low-buffered LAMP system designed for the conserved sequences of two genes, CARDS and gyrB, was applied. The rapid release of large amounts of H⁺ in the low-buffered LAMP solution led to a speedy increase in electrical signals captured by the ISFET array, eliminating the need for a sophisticated temperature cycling and optical system. The on-chip results showed that the device can accurately complete MP detection with a detection limit of about 10³ copies/mL (approximately 1 copy per reaction). In the final clinical validation, the detection results of eight throat swab samples using the ISFET sensors were fully consistent with the clinical laboratory diagnostic outcomes, confirming the accuracy and reliability of the ISFET sensors for use in clinical settings. And the entire process from sample lysis to result interpretation takes about 60 min. This platform has potential to be used for the point-of-care testing (POCT) of pathogen infections, providing a basis for the timely adjustment of diagnosis and treatment plans. Full article

The Investigator ESSplex SE QS Kit (Qiagen) is a next-generation polymerase chain reaction (PCR) kit that, in 60 min, amplifies 17 Short Tandem Repeat (STR) markers, including the five European Standard Set (ESS) loci (D10S1248, D12S391, D1S1656, D22S1045, D2S441), the SE33 marker, and the locus Amelogenin for sex determination. Two quality sensors (QS1 and QS2) are also co-amplified to check PCR performance. Since forensic laboratories carry out hundreds of DNA typings annually, we verified the kit’s performance using half reaction volumes with the aim of improving the number of samples that may be amplified with a single kit and consequently reducing laboratory costs. In the present study, intended as a technical note rather than internal validation, some control samples (oral swabs) with known DNA profiles and 40 real casework samples were analyzed. We observed that reducing the total reaction volume, while keeping all component ratios unaltered, yields DNA profiles comparable to those obtained using standard reaction volumes and with allele peaks higher than those with regular volumes. Using half volumes for PCR amplification enables the analysis of a larger number of samples compared to the standard protocol, thereby reducing laboratory costs without compromising the quality of the analysis. Full article

In this work, we present a compact LIBS sensor developed for characterization of samples on a crime scene following requirements of law enforcement agencies involved in the project. The sensor operates both in a tabletop mode, for aside measurements of swabbed materials or taken fragments, and in handheld mode where the sensor head is pointed directly on targets at the scene. The sensor head is connected via an umbilical to an instrument box that could be battery-powered and contains also a color camera for sample visualization, illumination LEDs, and pointing system for placing the target in focus. Here we describe the sensor’s architecture and functionalities, the optimization of the acquisition parameters, and the results of some LIBS measurements. On nano-plotted traces at silica wafer and in optimized conditions, for most of the elements the detection limits, in term of the absolute element masses, were found to be below 10 picograms. We also show results obtained on some representative materials, like fingerprints, swabbed soil and gunshot residue, varnishes on metal, and coated plastics. The last, solid samples were used to evaluate the depth profiling capabilities of the instrument, where the recognition of all four car paint layers was achieved. Full article

Strabismus can be caused by abnormal tension of the extraocular muscles (EOMs) attached to the eyeball in superior, inferior, lateral, medial, superior oblique, and inferior oblique positions. Evaluating the tension in each EOM is crucial for surgical planning in strabismus, which is conducted by adjusting the tension on the EOM. The purpose of this study was to develop a compact measuring device to non-invasively evaluate the active EOM tension. The proposed device employed a cotton-tipped medical swab to transfer the EOM tension connected to the force sensor as a non-invasive medium. The tilting angle of the swab and the force of active EOM tension were wirelessly transferred to a laptop computer for recording and real-time displaying of the measured values. The active EOM tensions for the four recti muscles were 101.7 ± 15.0 g (mean ± SD) for the lateral rectus; 88.0 ± 15.4 g for the medial rectus; 61.3 ± 6.8 g for the inferior rectus; and 121.3 ± 38.5 g for the superior rectus. These values were higher than the reported values of 45–60 g measured in previous studies. In the previous studies, however, the EOM was detached from the globe and attached to a strain gauge, and, thus, there were no passive elastic forces from ocular connective tissue, resulting in lower values compared with the current study. The previous methods were also complex and not suitable for clinical measurement. Thus, the proposed method, which is non-invasive and mimics the conventional force generation test with a cotton-tipped swab, could facilitate the evaluation of active EOM tension, both clinically in strabismus management and in research into understanding its pathophysiology. Full article

The SARS-CoV-2 pandemic has triggered many studies worldwide in the area of biosensors, leading to innovative approaches for the quantitative assessment of COVID-19. A nanostructured field-effect transistor (FET) is one type of device shown to be ultrasensitive for virus determination. FETs can be used as transducers to analyze changes in electrical current caused by the bonding of viral molecules to the surface of the semiconducting nanomaterial layer of the FETs. Although nano-transistors require simple setups amenable to be miniaturized for point-of-care diagnostic of COVID-19, this type of sensor usually has limited sensitivity in biological fluids. The reason behind this is the shortened screening length in the presence of high ionic strength solutions. In the frame of this study, we propose a methodology consisting of the FET surface modification with a hydrogel based on the star-shaped polyethylene glycol (starPEG), which hosts specific antibodies against SARS-CoV-2 spike protein in its porous structure. The deposition of the hydrogel increases the effective Debye length, preserving the biosensor’s sensitivity. We demonstrate the capability of silicon nanonet-based FETs to detect viral antigens and cultured viral particles in phosphate-buffered saline (PBS) as well as in human-purified saliva. Finally, we discriminated between positive and negative patients’ nasopharyngeal swab samples. Full article

The current status of microbiological testing methods for the determination of viable bacteria in complex sample matrices, such as food samples, is the focus of this review. Established methods for the enumeration of microorganisms, particularly, the ‘gold standard’ agar plating method for the determination of total aerobic viable counts (TVC), bioluminescent detection of total ATP, selective molecular methods (immunoassays, DNA/RNA amplification, sequencing) and instrumental methods (flow cytometry, Raman spectroscopy, mass spectrometry, calorimetry), are analyzed and compared with emerging oxygen sensor-based respirometry techniques. The basic principles of optical O₂ sensing and respirometry and the primary materials, detection modes and assay formats employed are described. The existing platforms for bacterial cell respirometry are then described, and examples of particular assays are provided, including the use of rapid TVC tests of food samples and swabs, the toxicological screening and profiling of cells and antimicrobial sterility testing. Overall, O₂ sensor-based respirometry and TVC assays have high application potential in the food industry and related areas. They detect viable bacteria via their growth and respiration; the assay is fast (time to result is 2–8 h and dependent on TVC load), operates with complex samples (crude homogenates of food samples) in a simple mix-and-measure format, has low set-up and instrumentation costs and is inexpensive and portable. 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

Recently, filter paper (FP)-based surface-enhanced Raman scattering (SERS) substrates have stimulated significant attention owing to their promising advantages such as being low-cost, easy to handle, and practically suitable for real-field applications in comparison to the solid-based substrates. Herein, a simple and versatile approach of laser-ablation in liquid for the fabrication of silver (Ag)-gold (Au) alloy nanoparticles (NPs). Next, the optimization of flexible base substrate (sandpaper, printing paper, and FP) and the FP the soaking time (5–60 min) was studied. Further, the optimized FP with 30 min-soaked SERS sensors were exploited to detect minuscule concentrations of pesticide (thiram-50 nM), dye (Nile blue-5 nM), and an explosive (RDX-1,3,5-Trinitroperhydro-1,3,5-triazine-100 nM) molecule. Interestingly, a prominent SERS effect was observed from the Au NPs exhibiting satisfactory reproducibility in the SERS signals over ~1 cm² area for all of the molecules inspected with enhancement factors of ~10⁵ and relative standard deviation values of <15%. Furthermore, traces of pesticide residues on the surface of a banana and RDX on the glass slide were swabbed with the optimized FP substrate and successfully recorded the SERS spectra using a portable Raman spectrometer. This signifies the great potential application of such low-cost, flexible substrates in the future real-life fields. Full article

An IoT-WiFi smart and portable electrochemical immunosensor for the quantification of SARS-CoV-2 spike protein was developed with integrated machine learning features. The immunoenzymatic sensor is based on the immobilization of monoclonal antibodies directed at the SARS-CoV-2 S1 subunit on Screen-Printed Electrodes functionalized with gold nanoparticles. The analytical protocol involves a single-step sample incubation. Immunosensor performance was validated in a viral transfer medium which is commonly used for the desorption of nasopharyngeal swabs. Remarkable specificity of the response was demonstrated by testing H1N1 Hemagglutinin from swine-origin influenza A virus and Spike Protein S1 from Middle East respiratory syndrome coronavirus. Machine learning was successfully used for data processing and analysis. Different support vector machine classifiers were evaluated, proving that algorithms affect the classifier accuracy. The test accuracy of the best classification model in terms of true positive/true negative sample classification was 97.3%. In addition, the ML algorithm can be easily integrated into cloud-based portable Wi-Fi devices. Finally, the immunosensor was successfully tested using a third generation replicating incompetent lentiviral vector pseudotyped with SARS-CoV-2 spike glycoprotein, thus proving the applicability of the immunosensor to whole virus detection. Full article

The detection of salivary cotinine is useful for convenient smoking tests in spite of the high background effect of saliva. For precise results, the conventional salivary cotinine analysis for smoking detection requires complex pretreatment processes. Hence, in this study, we developed a modified paper-based lateral flow immunoassay (LFIA), termed “gap-LFIA”, for the direct application of saliva collected using cotton swabs for on-site detection. The gap-LFIA was constructed by modifying a conventional LFIA sensor, where the sample pad was divided to have a 3 mm gap. A saliva-collected cotton swab was inserted into the gap, and then, a buffer solution was added to the outer sample pad to dilute the saliva automatically. The gap-LFIA reduced the interference in salivary samples and showed improved signals, allowing for using the whole saliva directly without additional steps. Further, the deviation of results using a strip was less than that when the saliva was not diluted in a conventional cotinine kit, and it helped to distinguish between smokers and non-smokers more clearly in 15 min. This method of automatic dilution may apply to various clinical samples, including blood and serum, for direct application in future detections. Full article

This paper discusses the application of two approaches (direct and inverse) to the identification of volatile substances by means of a gas sensor array in a headspace over nasal mucus swab samples taken from calves with differing degrees of respiratory damage. We propose a unique method to visualize sensor array data for quality analysis, based on the spectra of cross mass sensitivity parameters. The traditional method, which requires an initial sensor array trained on the vapors of the individual substances (database accumulation)—with their further identification in the analyzed bio-samples through the comparison of the analysis results to the database—has shown unsatisfactory performance. The proposed inverse approach is more informative for the pattern recognition of volatile substances in the headspace of mucus samples. The projection of the calculated parameters of the sensor array for individual substances in the principal component space, acquired while processing the sensor array output from nasal swab samples, has allowed us to divide animals into groups according to the clinical diagnosis of their lung condition (healthy respiratory system, bronchitis, or bronchopneumonia). The substances detected in the gas phase of the nasal swab samples (cyclohexanone, butanone-2,4-methyl-2-pentanone) were correlated with the clinical state of the animals, and were consistent with the reference data on disease markers in exhaled air established for destructive organism processes. Full article

Field-effect transistor (FET) biosensors have been intensively researched toward label-free biomolecule sensing for different disease screening applications. High sensitivity, incredible miniaturization capability, promising extremely low minimum limit of detection (LoD) at the molecular level, integration with complementary metal oxide semiconductor (CMOS) technology and last but not least label-free operation were amongst the predominant motives for highlighting these sensors in the biosensor community. Although there are various diseases targeted by FET sensors for detection, infectious diseases are still the most demanding sector that needs higher precision in detection and integration for the realization of the diagnosis at the point of care (PoC). The COVID-19 pandemic, nevertheless, was an example of the escalated situation in terms of worldwide desperate need for fast, specific and reliable home test PoC devices for the timely screening of huge numbers of people to restrict the disease from further spread. This need spawned a wave of innovative approaches for early detection of COVID-19 antibodies in human swab or blood amongst which the FET biosensing gained much more attention due to their extraordinary LoD down to femtomolar (fM) with the comparatively faster response time. As the FET sensors are promising novel PoC devices with application in early diagnosis of various diseases and especially infectious diseases, in this research, we have reviewed the recent progress on developing FET sensors for infectious diseases diagnosis accompanied with a thorough discussion on the structure of Chem/BioFET sensors and the readout circuitry for output signal processing. This approach would help engineers and biologists to gain enough knowledge to initiate their design for accelerated innovations in response to the need for more efficient management of infectious diseases like COVID-19. Full article

The rapid spread of the Coronavirus Disease 2019 (COVID-19) pandemic, caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pathogen has generated a huge international public health emergency. Currently the reference diagnostic technique for virus determination is Reverse Transcription Polymerase Chain Reaction (RT-PCR) real time analysis that requires specialized equipment, reagents and facilities and typically 3–4 h to perform. Thus, the realization of simple, low-cost, small-size, rapid and point-of-care diagnostics tests has become a global priority. In response to the current need for quick, highly sensitive and on-site detection of the SARS-CoV-2 virus in several aqueous solutions, a specific molecularly imprinted polymer (MIP) receptor has been designed, realized, and combined with an optical sensor. More specifically, the proof of concept of a SARS-CoV-2 sensor has been demonstrated by exploiting a plasmonic plastic optical fiber sensor coupled with a novel kind of synthetic MIP nano-layer, especially designed for the specific recognition of Subunit 1 of the SARS-CoV-2 Spike protein. First, we have tested the effectiveness of the developed MIP receptor to bind the Subunit 1 of the SARS-CoV-2 spike protein, then the results of preliminary tests on SARS-CoV-2 virions, performed on samples of nasopharyngeal (NP) swabs in universal transport medium (UTM) and physiological solution (0.9% NaCl), were compared with those obtained with RT-PCR. According to these preliminary results, the sensitivity of the proposed optical-chemical sensor proved to be higher than the RT-PCR one. Furthermore, a relatively fast response time (about 10 min) to the virus was obtained without the use of additional reagents. Full article

We propose a new telerobotic system for untact swab sampling to prevent the infection of medical staff during upper respiratory sample collection. The system consists of a slave robot and two master devices. The slave robot is designed to move a swab in 6 degrees of freedom within the facial area and to insert and remove the swab under remote control by an operator at the master site based on magnified imaging of the patient’s facial area. The insertion and removal of the swab into and from the nostril are implemented by means of a swab insertion unit; as the counterpart to this unit, the master system also includes a swab insertion device to control the swab insertion unit remotely. A force sensor installed on the swab holder enables monitoring of the force generated when the swab touches the target. In experiments, a virtual specimen was installed on the posterior nasopharynx wall of a life-size head phantom model. The nasopharyngeal swab samplings of the phantom model were successfully performed thanks to the force monitoring capability of the proposed telerobotic system, showing that this system is suitable for remote upper respiratory sample collection. Full article