Rapid Detection Method Research Articles

Comparative evaluation of phenotypic methods to detect methicillin resistance with mecA gene detection by PCR in Staphylococcus aureus

It is crucial to employ precise, reliable, and rapid detection methods to avoid the indiscriminate use of antimicrobial drugs and make informed decisions regarding appropriate treatment and the execution of efficient measures to prevent infections.: A group of 112 isolates from different clinical specimens, initially identified as using the cefoxitin disc diffusion test, underwent additional phenotypic tests such as the Oxacillin E strip and MRSA CHROM agar. These methods were then evaluated and compared with gene detection via PCR, which is regarded as the gold standard. Of the 112 isolates identified by the cefoxitin disc diffusion test, 101 (90.2%) tested positive for the gene. The Oxacillin E strip had a sensitivity of 98% and a specificity of 91%, while MRSA CHROM agar showed a sensitivity of 96.03% and a specificity of 82%. Among the 101 mec A-positive isolates, 44.5% met the CDC definition criteria for . : Our study concludes that phenotypic methods, including the cefoxitin disc diffusion test, are not completely reliable in detecting methicillin resistance in According to our results, combining the cefoxitin disc diffusion test with the oxacillin E strip is an effective approach for detecting in resource-constrained settings. Given the advantages of PCR, it is recommended to perform PCR for gene detection on a regular basis to identify strains in important clinical specimens.

Application of Aspergillus-specific immunoglobulin G antibody in the management of pulmonary aspergillosis

Aspergillus causes a range of diseases in patients with immunodeficiency or underlying pulmonary diseases. Aspergillus-specific immunoglobulin G antibody (Asp-IgG) is produced in response to Aspergillus pathopoiesis. Currently, Asp-IgG is included in the diagnostic criteria for chronic pulmonary aspergillosis (CPA) and allergic bronchopulmonary aspergillosis (ABPA). In recent years, some researches have proved that Asp-IgG also has important diagnostic value for non-neutropenic patients with invasive pulmonary aspergillosis (IPA). This review summarized the progress of Asp-IgG in the management of different types of pulmonary aspergillosis, and the advantages of rapid detection methods for Asp-IgG.

A novel single-tube LAMP-CRISPR/Cas12b method for rapid and visual detection of zoonotic Toxoplasma gondii in the environment

BackgroundToxoplasma gondii oocysts, excreted in cat feces, pose a significant health risk to humans through contaminated soil and water. Rapid and accurate detection of T. gondii in environmental samples is essential for public health protection.MethodsWe developed a novel, single-tube detection method that integrates loop-mediated isothermal amplification (LAMP), the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12b system, and lateral flow immunoassay strips for rapid, visual identification of T. gondii. This method targets the T. gondii B1 gene, initially amplifies it with LAMP, directed by a single-guide RNA (sgRNA). It then recognizes the amplified target gene and activates trans-cleavage, cutting nearby single-stranded DNA (ssDNA) reporters. Fluorescence detection was performed using a 6-Carboxyfluorescein (FAM)-12N-Black Hole Quencher-1 (BHQ1) reporter, while Fluorescein Isothiocyanate (FITC)-12N-Biotin enabled visual detection on lateral flow strips. The method was tested for its ability to detect various T. gondii genotypes and related parasites, assessing its specificity and broad-spectrum applicability. It was further applied to real-world environmental samples to evaluate its practicality.ResultsThe LAMP-CRISPR/Cas12b method exhibited high specificity and broad-spectrum detection capability, successfully identifying nine T. gondii genotypes and distinguishing them from 11 other parasitic species. Sensitivity testing at both molecular (plasmid) and practical (oocyst) levels showed detection limits of 10 copies/μL and 0.1 oocyst, respectively. When applied to 112 environmental samples (soil, water, and cat feces), the method demonstrated 100% sensitivity, accurately reflecting known infection rates.ConclusionsThis LAMP-CRISPR/Cas12b single-tube method offers a robust, innovative approach for monitoring zoonotic T. gondii in environmental samples, with significant implications for public health surveillance.Graphical

Establishment and application of a rapid new detection method for antimicrobial susceptibility testing of Klebsiella pneumoniae based on MALDI-TOF MS.

The earlier appropriate treatment of Klebsiella pneumoniae infections according to the antimicrobial susceptibility profile based on the minimum inhibitory concentration (MIC) has a great clinical benefit. Our objective was to establish a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based antimicrobial susceptibility test (AST) for K. pneumoniae that produces reliable results within a few hours. Our rapid method is similar to the classical turbidity-based microdilution method. We confirmed, theoretically and experimentally, that MALDI-TOF MS can replace the naked eye in judging the results (MICMS) by "seeing" the bacterial growth in the presence of different concentrations of antibiotics, including determination of the lower limit of bacterial count detection (4 × 105 cfu), the optimal period of incubation (2 h), and bacterial growth curve assay. Based on the study mentioned above, we determined the susceptibility of K. pneumoniae to imipenem. The MICMS and MIC data agreed over 85% (40/46) within 1 dilution range. Susceptibility profiles determined with our rapid method and the reference broth microdilution method were also compared. MICMS resulted in 97.9% (45/46) category agreement, 2.2% minor discrepancies, no major discrepancies, and no very major discrepancies. The summarized category agreement resulted in a kappa coefficient of almost 1 for weighted Cohen's kappa, which could be considered a nearly perfect agreement. It took just 2 h to produce a susceptibility profile with a low failure rate using our new rapid AST method, a work day earlier than the broth microdilution method.IMPORTANCEEmpirical antimicrobial use before antimicrobial susceptibility test (AST) is necessary but risks patient harm and excess costs. It is particularly worrying that the inappropriate use of carbapenems has allowed carbapenem-resistant Klebsiella pneumoniae to become the commonest transmissible carbapenem-resistant Enterobacterales worldwide. Guidelines recommend targeted therapy based on minimum inhibitory concentration results, which directly reflect the effectiveness of antibacterial drugs. The gold standard method of AST relies on visible bacterial growth, causing long turnaround times. Current rapid AST techniques are hampered by factors such as high costs, technological complexities, and limited detection capabilities. We present a novel rapid method and applied to the determination of the susceptibility of K. pneumoniae to imipenem. It took just 2 h to produce a susceptibility profile with a low failure rate, a work day earlier than the standard method. Our method is potentially a faster, more precise, cost-efficient, and user-friendly AST method that can enhance the effectiveness of treatment strategies.

Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection

In this study, two methods for assessing the purity of A2 fermented milk and β-casein genotypes in dairy cows were examined. The need for rapid and precise methods for herd screening and A2 milk quality control justified this study. Accordingly, an ELISA test was developed to identify β-casein A1 in fermented milk, and a commercial Lateral Flow Immunoassay (LFIA) was evaluated to determine A2A2 genotypes and the purity of A2 milk. The results demonstrated 100% sensitivity and specificity of the ELISA test in identifying β-casein A1. The LFIA test successfully identified A2A2 genotypes and confirmed the purity of A2 milk, with a minimum detectable contamination of 5% for raw milk and 10% for fermented milk. Both tests exhibited 100% sensitivity and specificity, resulting in positive and negative predictive values of 100%. The positive likelihood ratio was infinite, while the negative was zero, indicating a precise and reliable test with no false diagnoses. Compared to traditional genotyping, these methods proved to be more practical and showed potential for large-scale screening. It was concluded that ELISA and LFIA are valuable tools for ensuring the quality and authenticity of A2 milk, meeting the demands of producers and consumers for safe and healthy dairy products.

A rapid and high-throughput fraud detection method of Philippine coconut wine (lambanog) using 1H qNMR spectroscopy

Methanol contamination of the Philippine coconut spirit lambanog (often called coconut wine) is the major cause of lambanog-related deaths in the country. Hence, a strict quality control and detection method must be established for methanol in tandem with ethanol analysis. In this study, a quantitative Nuclear Magnetic Resonance spectroscopy (qNMR) method using 1H analysis was developed to quantify the methanol and ethanol in 26 lambanog samples collected from four different provinces in Luzon, Philippines. A certified qNMR standard was used as an internal standard for the 1H NMR analyses to increase the accuracy of the measurements. The calculated limit of detection and limit of quantification of methanol with values of 0.076%(v/v) and 0.25%(v/v), respectively, were sufficiently low and allow the monitoring of methanol within the acceptable safety level. Moreover, the results of methanol and ethanol analysis using the proposed method were in good agreement with those obtained from GC-FID analysis which is the conventional method for alcohol analysis. In contrast to GC-FID, the qNMR method for simultaneous alcohol analysis can provide results in a shorter period. The results of this study show the potential of the qNMR method to be used as an alternative analytical method which widens the range of analytical possibilities to detect methanol in lambanog samples for the safety of the consumers.

A rapid liquid chromatography-tandem mass spectrometry based method for the detection of Tet(X) resistance gene in Enterobacteriaceae

There is a major public health threat posed by antibiotic resistance around the world. Tigecycline overcomes the resistance mechanisms of traditional tetracyclines and is often seen as the final resort in combating infections caused by bacteria resistant to multiple drugs. However, the introduction of new mobile tet(X) tetracycline destructases is leading to a notable rise in tigecycline resistance. Therefore, a rapid detection method is needed to monitor the spread of tigecycline resistance. In this study, a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to detect tet(X) in bacterial isolates was developed. This method utilized the analysis by LC-MS/MS of metabolite ratios to determine the presence of tet(X). Bacterial suspensions were co-incubated with tigecycline for 1 h, where tet(X) destructase inactivated tigecycline, making a particular metabolite with a 16-Da change in mass. The characterized quantitative ion pairing of tigecycline in the ESI positive mode was observed at 586.1 → 569.1 m/z. The oxygenated tigecycline detection was established at 602.2 → 529.1 m/z. A model was established using 35 tet(X)-positive and 15 tet(X)-negative Enterobacteriaceae strains in this study to optimize the cutoff value. Applying the model to analyze 70 bacterial isolates, the sensitivity of the LC-MS/MS test was 98.9% compared to polymerase chain reaction (PCR), and specificity was 100%. This method is rapid and easy to operate, providing results within 1 h, making it more suitable for routine use in clinical microbiology laboratories.

Development of a Colloidal Gold Immunochromatographic Strip for Specific Detection of Photobacterium damselae subsp. piscicida.

Photobacterium damselae subsp. piscicida (Pdp) is characterised by high infectivity and mortality and wide distribution, resulting in substantial economic losses to the global aquaculture industry. Therefore, it is critical to develop a simple, rapid and accurate diagnostic method for Pdp detection. Herein, two Pdp-specific monoclonal antibodies (mAbs 1H7 and 6E6) were obtained, indirect enzyme-linked immunosorbent assay and immunofluorescence analysis confirmed that the mAbs specifically recognised the outer membrane of Pdp but could not recognise the other Gram-positive or Gram-negative bacteria, indicating good specificity. A colloidal gold immunochromatographic strip (CGICS) was developed here for Pdp detection by optimising the colloidal gold labeling and antibody coating conditions using 1H7 as a colloidal gold-labelled antibody and 6E6 as a coated antibody. The CGICS exhibited a detection limit of 1 × 105 CFU/mL for Pdp and a detection time of 10 min. Cross-reaction analysis revealed that the CGICS could specifically detect different Pdp isolates without cross-reaction with the other bacterial species or the subspecies P. damselae subsp. damselae. Stability analysis revealed that the CGICS were stored in closed plastic packaging at 25°C and 4°C for more than 8 months. The CGICS retained their detection limit after 6 months at 4°C, and 4 months at 25°C. Using tissues from the experimentally challenged Lateolabrax maculatus, the CGICS and polymerase chain reaction revealed a 100% coincidence rate. These findings indicated that the established CGICS exhibits high specificity and sensitivity. It is rapid and simple for Pdp detection, making it a potential on-site tool for Pdp detection in fish aquaculture.

Rapid detection of carbapenemase production in Aeromonas using phenotypic tests based on colorimetric microtube assay.

Antibiotic resistance, particularly carbapenem resistance, poses a significant global health threat due to the limited availability of effective antibiotics. Carbapenem-resistant Aeromonas are increasingly recognized for their role in various infections, necessitating rapid and accurate detection methods. This study aimed to evaluate several phenotypic tests, including the Carba NP test (CNPt), Carba NP-direct test (CNPd), and Blue-Carba test (BCT), for their effectiveness in rapidly detecting carbapenemase production in Aeromonas. These tests target both the chromosomally encoded CphA metallo-β-lactamase (MBL) and acquired carbapenemases. Additionally, a modified phenotypic test called the Colony-Carba NP test (c-CNPt) was introduced to enhance sensitivity and specificity. A retrospective analysis was conducted on 131 clinically conserved Aeromonas strains harboring identified carbapenem resistance genes, using CNPt, CNPd, BCT, and the newly developed c-CNPt and EDTA-Colony-Carba NP test (ec-CNPt). The stability of c-CNPt reagents stored at -80°C was also assessed. Additionally, a prospective study conducted from July 2021 to November 2023 evaluated 152 Aeromonas isolates to determine the clinical applicability of these tests. Our results demonstrated that CNPd and BCT achieved 100% sensitivity and specificity, surpassing the traditional CNPt, which showed only 63.6% sensitivity for Aeromonas strains. The c-CNPt also showed 100% sensitivity and specificity, with the ec-CNPt effectively differentiating between MBL and serine carbapenemase types. Stability tests confirmed that c-CNPt reagents could be stored at -80℃ for up to 1 year without performance degradation. These findings highlight the practicality and reliability of these phenotypic tests for routine laboratory use, providing a rapid and cost-effective method for detecting carbapenemase production.The rapid detection of carbapenemase production in Aeromonas is of paramount importance due to the significant clinical and public health implications associated with antibiotic resistance. The development and validation of rapid phenotypic tests such as the Colony-Carba NP test (c-CNPt) and the EDTA-Colony-Carba NP test (ec-CNPt) are crucial advancements in the field. These tests offer a highly sensitive and specific method for detecting carbapenemase production in Aeromonas, including the differentiation between metallo-β-lactamase and serine carbapenemases. The c-CNPt and ec-CNPt are cost-effective, easy to perform, and provide rapid results, making them suitable for routine clinical use. Additionally, the stability of the reagents ensures their practicality for long-term application in various healthcare settings. Implementing these phenotypic tests in clinical laboratories can significantly enhance the early detection and appropriate treatment of carbapenem-resistant Aeromonas infections.

Point-of-care testing diagnosis of African swine fever virus by targeting multiple genes with enzymatic recombinase amplification and CRISPR/Cas12a System

African swine fever virus (ASFV) infection is causing devastating outbreaks globally; pig farming has suffered severe economic losses due to the ASFV. Currently, strict biosecurity control measures can mitigate the incidence of ASF. Rapid, cost-effective, and sensitive detection of ASFV can significantly reduce disease transmission and mortality. CRISPR/Cas-associated proteins can detect polymorphisms with high specificity and sensitivity, making them ideal for detecting pathogens. In this study, based on CRISPR/Cas12a integrated with enzymatic recombinase amplification (ERA) technology, a CRISPR/Cas12a detection system capable of identifying ASFV E183L, K205R, and C962R gene sequences has been developed. The ERA-CRISPR/Cas12a detection system detected ASFV precisely without cross-reactivity with other porcine pathogen templates and with a sensitivity detection limit of 10 copies per reaction; it takes 60 minutes to complete the detection process. In combination with this integrated ERA pre-amplification and Cas12a/crRNA cutting assay, it provides a rapid, straightforward, sensitive, and specific method for ASFV detection in the field.

A New Method for Rapid Detection of Surface Defects on Complex Textured Tiles

A New Method for Rapid Detection of Surface Defects on Complex Textured Tiles

Advances in the detection technology of vegetable soil borne fungi and bacteria

Soil borne diseases are one of the most serious diseases which often results the decline of vegetables quality and loss of production. Moreover, it is difficult for plants to exhibit disease symptoms in the early stages attributing to strong concealment of soil borne pathogens. Therefore, early detection of pathogens and their physiological races plays an important role in reducing the harm of pathogens associated with diseases of vegetable crops. The traditional diagnostic techniques relied on the time consuming and less accurate methods like disease symptom observation, microscopic diagnosis, and culture techniques etc. The development of molecular biology technology has brought revolutionary changes to the diagnosis of vegetable soil borne diseases, improving the accuracy and efficiency of diagnosis. This paper reviews the various molecular detection techniques for vegetable soil borne pathogens (PCR, nested-PCR, multiplex PCR, etc.) and their physiological races (host identification, DNA molecular markers, transposon detection, etc.), explains the advantages and disadvantages of each detection technique. Furthermore, the paper comprehensively introduces the application of molecular detection technology for soil borne pathogen detection in soil, plants, and seeds. Finally, we put forward important perspectives for the future development of rapid detection methods, aiming to promote rapid diagnosis of soil pathogenic microorganisms and provide guidance for the control of biological risks.

FastQAFPN-YOLOv8s-Based Method for Rapid and Lightweight Detection of Walnut Unseparated Material

Walnuts possess significant nutritional and economic value. Fast and accurate sorting of shells and kernels will enhance the efficiency of automated production. Therefore, we propose a FastQAFPN-YOLOv8s object detection network to achieve rapid and precise detection of unsorted materials. The method uses lightweight Pconv (Partial Convolution) operators to build the FasterNextBlock structure, which serves as the backbone feature extractor for the Fasternet feature extraction network. The ECIoU loss function, combining EIoU (Efficient-IoU) and CIoU (Complete-IoU), speeds up the adjustment of the prediction frame and the network regression. In the Neck section of the network, the QAFPN feature fusion extraction network is proposed to replace the PAN-FPN (Path Aggregation Network—Feature Pyramid Network) in YOLOv8s with a Rep-PAN structure based on the QARepNext reparameterization framework for feature fusion extraction to strike a balance between network performance and inference speed. To validate the method, we built a three-axis mobile sorting device and created a dataset of 3000 images of walnuts after shell removal for experiments. The results show that the improved network contains 6071008 parameters, a training time of 2.49 h, a model size of 12.3 MB, an mAP (Mean Average Precision) of 94.5%, and a frame rate of 52.1 FPS. Compared with the original model, the number of parameters decreased by 45.5%, with training time reduced by 32.7%, the model size shrunk by 45.3%, and frame rate improved by 40.8%. However, some accuracy is sacrificed due to the lightweight design, resulting in a 1.2% decrease in mAP. The network reduces the model size by 59.7 MB and 23.9 MB compared to YOLOv7 and YOLOv6, respectively, and improves the frame rate by 15.67 fps and 22.55 fps, respectively. The average confidence and mAP show minimal changes compared to YOLOv7 and improved by 4.2% and 2.4% compared to YOLOv6, respectively. The FastQAFPN-YOLOv8s detection method effectively reduces model size while maintaining recognition accuracy.

Diagnostic potential of rhinovirus C using reverse-transcription loop-mediated isothermal amplification (RT-LAMP).

Rhinoviruses (RVs), particularly RV-C, frequently cause acute respiratory infections and asthma exacerbations. However, there is a lack of routine detection methods. Thus, this study aims to develop a rapid molecular and differential diagnostic detection method for RV-C using the reverse transcription (RT) loop-mediated isothermal amplification (LAMP) approach. The RT-LAMP assay targeting the 5'UTR region of RV-C genome was optimized by varying the reaction temperature, magnesium sulphate, betaine concentrations, and reaction time. Compared with conventional RT-PCR with a sensitivity of 106 copies of RNA, RT-LAMP demonstrated a significant increase in efficiency and sensitivity with a quantifiable viral load of at least 101 copies of RNA by gel electrophoresis and colour change, and 104 copies of RNA for end-point detection with a turbidimeter for 40min. The assay is also specific without amplifying RV-A16 and RV-B72 genomic RNA. In the proof-of-concept assay using 30 clinical respiratory samples with known etiological agents, it detected all RV-C isolates, of which its accuracy was confirmed by sequencing. The newly developed RT-LAMP assay demonstrated good analytical sensitivity and specificity toward RV-C. The assay provides an alternative for improved RV-C diagnosis.

Development and validation of a portable device for lab-free versatile nucleic acid extraction.

Nucleic acid testing (NAT) has revolutionized diagnostics by providing precise, rapid, and scalable detection methods for diverse biological samples. These recent advancements satisfy the increasing demand for on-site diagnostics, yet sample preparation remains a significant bottleneck for achieving highly sensitive diagnostic assays. There is an unmet need for compatible, efficient, and lab-free sample preparation for point-of-care NAT. To address this, we developed a portable, lab-free, and battery-powered device for extracting nucleic acids. We explored using low centrifugal forces with existing commercial chemistry, demonstrating excellent performance. We designed and tested a battery-powered device to enable lab-free extractions, and verified reagents stored out to 6 months, suggesting exceptional deployment capabilities. We evaluated our device, comparing our results against those from a benchtop centrifuge across three types of samples: HIV RNA in buffer, HIV RNA in plasma, and SARS-CoV-2 RNA in saliva. The portable device demonstrated excellent agreement with the benchtop centrifuge, indicating high reliability. By providing an effective on-site sample preparation solution, the widespread adoption of low centrifugal extractions will improve the sensitivity and reliability of NAT and will positively impact other point-of-care technologies such as next generation sequencing (NGS), biomarker detection, and environmental monitoring.

In vitro/In vivo oxygen electrochemical nanosensor for bioanalysis

Direct in vivo monitoring of O2 is critical in the study of numerous biological processes, and the development of novel highly sensitive techniques for O2in vivo detection is of great potential importance. Electrochemical amperometric sensors are some of the most promising, easy to operate, inexpensive, sensitive and have a high temporal resolution. As part of this work, we have demonstrated a direct rapid method for molecular oxygen detection inside multicellular spheroids in vitro and rat brain in vivo. External and internal oxygen profiles in human adenocarcinoma MCF-7 spheroids were studied using developed nanoelectrode. The size of spheroids was shown to affect the level of hypoxia inside them, and also affected the oxygen consumption near spheroids in solution. To demonstrate the in vivo relevance of the novel sensor we used it to measure the oxygen concentrations in the superficial layers of the rat brain. This study demonstrates a minimally invasive electrochemical method for real-time oxygen profiling in vivo.

Using diatom chain length as a bioindicator of heavy-metals contamination in marine environments

The increasing release of toxic heavy metals into marine environments poses significant risks due to their persistence and bioaccumulation. Diatoms are ideal bioindicators because of their sensitivity to environmental changes. Despite traditional methods for detecting these persistent pollutants effectively identify composition and concentration, they are time-consuming, they often require the use of harmful reagents, and do not allow a fast assessment of detrimental impacts on marine organisms. To fill this gap, we have successfully investigated the toxicity of different heavy metals in the marine diatom Skeletonema pseudocostatum thanks to a newly developed high-power terahertz (THz) spectrometer. By combining THz spectroscopy, microscopy and ecotoxicological assays, we found that the formation of long diatom chains is significantly inhibited by the presence of lead, copper, and chromium, which disrupt their metabolism. Although the THz absorption and refractive index spectra were not affected by diatom concentration in undoped samples, THz frequencies were highly sensitive to changes in diatom chain length due to heavy metals exposure. These findings suggest that this approach allows to investigate the biochemical processes involved in chain formation in S. pseudocostatum and related algae. THz spectroscopy could therefore provide deeper insights into the microscopic metabolic activity of diatoms, addressing key biochemical questions surrounding these organisms. Furthermore, we propose this novel approach for environmental pollution monitoring, since it could provide a rapid, harmless and sensitive detection method to assess heavy metal toxicity in marine diatoms, key organisms at the basis of the trophic chain.

Rapid and multiple visual detection of Fasciola hepatica in feces via recombinase polymerase amplification integrated with CRISPR/Cas12a technology

Fasciola hepatica is a foodborne zoonotic parasite causing significant economic losses and impacting human and livestock health in resource-limited regions. We developed a rapid, reliable, and sensitive detection method combining recombinase polymerase amplification (RPA) with CRISPR/Cas12a, allowing visualization with the naked eye or a fluorescence reader. Multiple visual methods were used to analyze the assay results. Fluorescence signals were collected using a fluorescence reader or observed under UV or blue light. Lateral flow strips (LFS) were used for visual detection. Among seven primer pairs and three CRISPR RNA (crRNA) screened, F1/R1 and crRNA3 were optimal. The Cas12a reaction buffer was optimized with 50 mM Tris-HCl and 80 mM NaCl, with an RPA reaction time of 20 min. The assay showed high specificity and excellent sensitivity for F. hepatica, detecting 0.122 copies/μL with fluorescence and 8.6 copies/μL with LFS. Testing of 143 sheep and 43 human fecal samples showed 98.39 % consistency with qPCR and nested PCR, with prevalence rates of 52.45 % and 18.6 % in sheep and humans, respectively. Our assay offers substantial potential for point-of-care testing in resource-limited areas, addressing the need for rapid and accurate diagnosis of F. hepatica.

One stone two birds: NH2-UiO-66@MB-based bimodal aptasensor for sensitive and rapid detection of zearalenone in cereal products

Zearalenone (ZEN), a prevalent mycotoxin found in cereal crops, poses a significant threat to food safety and human health. To address this issue, there is an urgent need for rapid, sensitive, and cost-effective detection methods. In this study, we developed a novel bimodal aptasensor based on NH2-UiO-66@MB composites for ZEN detection. Methylene blue (MB) was encapsulated within NH2-UiO-66 nanoparticles to enhance fluorescence intensity, and the ZEN aptamer was attached to NH2-UiO-66 via Zr-O-P bonds and Coulombic forces to create a gating effect. In the presence of ZEN, the aptamer bounded to ZEN, triggering a conformational change that released MB, resulting in decreased fluorescence intensity and absorbance. Under optimal conditions, the bimodal aptasensor exhibited wide linear ranges (2.5-300 ng/mL for colorimetry and 0.125-200 ng/mL for fluorescence) and low limits of detection (1.16 ng/mL for colorimetry and 0.03 ng/mL for fluorescence). Furthermore, the aptasensor demonstrated high specificity and reproducibility, with recovery rates ranging from 97.43% to 103.71% in spiked cereal products. These findings highlight the potential of the developed bimodal aptasensor as a novel tool for the rapid, accurate, and cost-effective detection of ZEN in cereal products, contributing to improved food safety monitoring.

An OPRM1-SNAP-tag/CMC method to directly identify drug components in sewage.

The scourge of drug addiction and abuse poses a significant challenge to society. Opioid drugs acting on μ-opioid receptor (OPRM1) make it one of the pivotal targets for drug addiction. In the past decade, sewage analysis has become a prevalent method of drug monitoring. However, traditional methods of detecting drugs in sewage are cumbersome, and rapid detection methods are relatively lacking. To address this, an innovative OPRM1-SNAP-tag/CMC method to directly identify drug components in sewage was established. Cell membrane chromatography (CMC) is an affinity chromatography technique which effectively detects receptor affinity substances. Cells constructed with high expression of specific receptor could be used to screen for compounds acting on the receptor. CMC based on OPRM1 provides a potentially convenient and effective tool for the detection of targeted drug components in sewage. In this study, the selectivity, reproducibility, column lifetime, and carryover of the CMC column had been assessed. Initially, we eluted the collected domestic sewage with methanol and acetonitrile, and the retention peaks were observed on the CMC system. Subsequently, without any preliminary sample preparation, we directly injected filtered samples of suspicious sewage into the OPRM1-SNAP-tag/CMC system, where we observed retention peaks as well. The retained components were further identified as morphine by using UPLC-MS/MS. In conclusion, the OPRM1-SNAP-tag/CMC method stands out as a reliable and robust model for the detection of drug components in sewage. It provides a valuable analytical tool for frontline drug control efforts, enhancing our capacity to monitor and mitigate the impact of drug abuse on society.