Rapid Detection Method Research Articles

CRISPR/Cas13a-mediated visual detection: A rapid and robust method for early detection of Nosema bombycis in silkworms

The sericulture industry faces a significant threat from the Pebrine disease of silkworms, caused by Nosema bombycis. Nonetheless, the current microscopic diagnostic methods can be time-consuming, labor-intensive, and lacking sensitivity and accuracy. Therefore, it is crucial to develop a novel detection approach that is efficient, highly sensitive, and low-cost. In this regard, the CRISPR/Cas system has the potential to be a fast, accurate, and highly specific method of detection. Herein, using a microplate reader, a portable fluorescence detection device, and test strips as signal output tools respectively, we have efficiently developed three rapid and facile visual detection methods for N. bombycis using a CRISPR/Cas13a system with conjugation of Recombinase polymerase amplification (RPA). We evaluated the sensitivity of this combined technology by comparing it with the positive plasmid standard and the genome standard of N. bombycis. Remarkably, the sensitivity of the CRISPR/Cas13a system for N. bombycis positive plasmid standard based on the microplate reader, portable fluorescence detection device, and test strips was 1 copy/μL, 10 copies/μL, and 1 copy/μL, respectively, while for the N. bombycis genome standards, the detection sensitivity was 10 fg/μL, 10 fg/μL, and 1 fg/μL, respectively. In addition, extensive evaluations have demonstrated that the established technology can accurately detect N. bombycis without cross-reactivity with other pathogens, ensuring a specificity rate of 100%. In brief, this study will provide a practical, efficient, and affordable method for early and rapid detection of N. bombycis in various settings.

Circular DNA enhanced amplification-free CRISPR/Cas12a assays for end-user friendly ultra-sensitive Porphyromonas gingivalis diagnosis

Periodontitis not only leads to tooth loss but is also associated with various systemic symptoms. Porphyromonas gingivalis (Pg) is the microorganism most closely linked to periodontitis. A rapid and accurate Pg detection method holds significant potential health benefits for patients. While several nucleic acid amplification-based diagnostic methods have been developed, there is a current lack of rapid diagnostic tools for Pg, particularly at chair-side level. Here, two detection methods for the Pg 16S gene based on the CRISPR/Cas12a system have been developed. To achieve ultra-high sensitivity in the developed CRISPR/Cas12a assays without pre-amplification, we designed a circular DNA molecule (termed as Cir-amplifier) which amplifies the signal output of the Cas12a reactions in a cascade response manner. The Cir-amplifier enhanced Fluorescence-Cas12a reaction achieved a minimum detectable limit of 0.39 CFU of Pg within 80-minute under room-temperature. In addition, the Cir-amplifier enhanced Lateral Flow Detection (LFD)-Cas12a reaction has demonstrated the ability to detect a minimum of 3.9 CFU of Pg in 90 min, with results observable by the naked eye. The potential of the two methods for Pg detection in clinical samples has been demonstrated, showing 100 % sensitivity compared to the traditional real-time PCR method. In summary, we have established two efficient detection methods for Pg that eliminate the need for pre-amplification, hence no amplicon contaminations, making them promising candidates for chair-side Pg diagnosis. Furthermore, the Cir-amplifier has been demonstrated to be adaptable to CRISPR/Cas systems producing various signal formats. The Cir-amplifier is expected to become a universal CRISPR/Cas enhancer.

Development and optimization of the qPCR-based rapid detection method for Chinese species of Sanghuangporus

Sanghuangporus, well known as ‘Sanghuang’, is a macrofungal genus accommodating 11 edible and medicinal species in China. With the prosperous development of ‘Sanghuang’ business in China, the fraudulent practice naming the products of other morphologically and phylogenetically related species as ‘Sanghuang’ happens frequently, which disturbs the market order of ‘Sanghuang’ business. To promote the healthy industrial development of Sanghuangporus and also prevent the loss of this macrofungal resource, we develop a qPCR-based rapid detection method in this study. Internal transcribed spacer sequences of Fuscoporia, Inonotus, Phellinus, Sanghuangporus and Tropicoporus were aligned to design Sanghuangporus-specific amplification primers and probe, which can specifically distinguish all 11 Chinese species of Sanghuangporus from species of other closely related genera. The concentrations of Sanghuangporus-specific primers and probe were optimized as 0.7 μmol/L and 0.5 μmol/L, respectively, where the qPCR has the minimum detectable DNA template concentration of 10−3 ng/μL. In summary, the current qPCR-based detection method was specific, sensitive and rapid for Chinese species of Sanghuangporus.

Single-atom nanozymes possessing robust multienzyme-mimetic catalytic properties for sensing of volatile alkaline gas

Volatile alkaline gases (VAGs) emitted from various sources, such as the food or chemical industry, pose potential harm to the natural environment and human health. Therefore, the development of a real-time, rapid, and cost-effective detection method is crucial. In this study, the enzyme-like catalytic types and activities of four different metal-based single-atom enzymes (Ga, Cu, Mn, and Zn SAzymes) were selected and evaluated. Among them, Ga SAzyme exhibited the most promising catalytic properties. Furthermore, the multienzyme catalytic properties of Ga SAzyme were utilized for the detection of ammonia, and it was found that its peroxidase-like (POD-like) activity showed a strong affinity for ammonia (Km = 0.05 mM). This detection strategy demonstrated high sensitivity, with a limit of detection (LOD) of 3.0 mM in the linear range of 0.01–0.05 M and 7.0 mM in the linear range of 0.075–0.25 M. Additionally, the method was characterized by fast response time (15 s) and low cost ($0.035 per sample). The proposed method holds great potential for the detection of VAGs from various sources in the future.

Detection of carbapenem-resistant gram-negative bacilli in Japan using the fully automated bacterial testing device RAISUS S4

ObjectiveWe investigated a rapid detection method for carbapenemase-producing gram-negative bacilli (CP-GNR) using meropenem (MEPM) to assess the efficiency of the antimicrobial susceptibility testing. MethodsWe used the function that can monitor the growth curve with the resistant bacteria monitoring function (RAISUS S4). Rapid detection of CP-GNR was performed using RAISUS S4 in two types of antimicrobial susceptibility testing, the RAISUS 18-hour method (18-h method) and RAISUS rapid method (rapid method) for Enterobacterales (F-GNR) and non-fermenting Gram-negative bacilli (NF-GNR). ResultsWhen F-GNR were based on MEPM MIC ≥ 0.25 μg/mL, CP-GNR were detected with a sensitivity of 100% (58/58) for the 18-h method and 98.3% (57/58) for the rapid method; the shortest detection times were 5.3 and 4.0 h, respectively. When NF-GNR were based on MEPM MIC > 8 μg/mL, it was possible to detect CP-GNR with 100% sensitivity (58/58) in both methods. Furthermore, in the analysis using the 18-h method for monitoring resistant bacteria, when ≥ 2 µg/mL was used as the screening concentration for F-GNR, approximately 50% of the resistant genotypes, NDM, GES, and KPC, were detected in approximately 7 h. However, detecting the IMP and VIM took 11–12 h. ConclusionsThe 18-h and rapid methods with RAISUS S4 were highly correlated with the results of the microdilution method of CLSI, and CP-GNR detection was rapid using a function that can monitor the growth curve with RAISUS S4.

Enzyme-free immunoassay for rapid, sensitive, and selective detection of C-reactive protein.

C-reactive protein (CRP) is a protein made by the liver, which is released into the bloodstream in response to inflammation. Furthermore, CRP is a potential risk factor for heart disease. Hence, it is of great importance to develop a rapid, sensitive, accurate, and cost-effective method for CRP detection. Herein, we report an enzyme-free fluorescent assay for the rapid and ultra-sensitive detection of CRP with a limit of detection (LOD) reaching as low as 3.08 pg/mL (i.e., ~ 27 fM). The high sensitivity of our method was simply achieved via dual-functionalized gold nanoparticles (AuNPs). By regulating the molar ratio of DNA to CRP antibody immobilized on the AuNP surface, hundreds to thousands-fold amplification in the analyte signal could be instantly accomplished. Furthermore, our sensor was selective: non-target proteins such as interleukin-6, interleukin-1β, procalcitonin, bovine serum albumin, and human serum albumin did not interfere with the target CRP detection. Moreover, simulated serum samples were successfully analyzed. Given the excellent sensitivity, selectivity, and high resistance to complicated matrices, the enzyme-free CRP detection strategy developed in this work can be used as a generic platform to construct sensors for a wide variety of protein biomarkers and hence offers potential as a tool for rapid, accurate, and low-cost medical diagnosis.

Development of an on-site real-time dual detection method for norovirus and rotavirus using RPA-CRISPR/Cas12,13

Norovirus (NoV) and rotavirus (RoV) are the two most prevalent foodborne viruses responsible for infectious gastroenteritis, with high contamination rates in fruits and vegetables. Monitoring and accurately identifying these pathogens are crucial for preventing human foodborne illnesses. Therefore, developing a rapid, highly specific, and sensitive detection method for NoV and RoV is essential. An on-site real time dual detection method for Noroviruses of genogroup I (GI-NoV) and Rotavirus of group A (A-RoV) was established by integrating Recombinase Polymerase Amplification (RPA) with the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated protein (Cas) system. The DNA amplification products generated by multiplex RPA were simultaneously detected by Cas12a and Cas13a in a single tube. Utilizing the trans-cleavage preferences of Cas12a and Cas13a, the specific cleavage of DNA and RNA probes carrying different fluorescent groups within the reaction system was achieved. The results indicated that the established dual detection method had a detection limit of 102 copies/μL for GI-NoV and A-RoV. Specificity assay showed the established method only detected GI-NoV and A-RoV, with no cross-reactivity with other four foodborne viruses. Furthermore, the detection capability of this method was validated in strawberries and lettuce. The method enables the simultaneous detection of GI-NoV and A-RoV in fresh produce at 37 °C within 60 min. Detection results were interpreted by measuring fluorescence values or observing color changes under blue light at the end of the reaction. This technique required minimal instrumentation, provided excellent visual representation of results, and served as a technical reference for the rapid on-site detection of foodborne viruses in food matrices.

Blood-rsCDM: a new rapid and simplified carbapenemase detection method for detecting carbapenemases in Enterobacterales directly from positive blood cultures

ObjectiveWe aim to validate and evaluate a new rapid and simplified method, called Blood-rsCDM, for the detection and characterization of carbapenemase using 3-aminophenylboronic acid (APBA) and ethylenediaminetetraacetic acid (EDTA) β-lactamase inhibitors from positive blood cultures.MethodWe utilized a panel of 172 Enterobacterales strains, including blaKPC (77), blaNDM (48), blaIMP (9), blaVIM (2), blaOXA-181 (2), blaKPC and blaNDM (6), as well as 28 carbapenem-susceptible Enterobacterales isolates, to assess the performance of Blood-rsCDM and the EDTA-carbapenem inactivation method (eCIM). Carbapenemase class was determined using specific inhibitors at 4 h and 6 h by Blood-rsCDM.ResultsBlood-rsCDM exhibited a sensitivity of 97.9% at both time points, with a specificity of 100%, regardless of the culture duration. The sensitivity of eCIM was 94.4%, with a specificity of 100%. Blood-rsCDM accurately characterized KPC-producing isolates as 77/77, metallo-β-lactamases (MBLs) as 58/59, and KPC and NDM carbapenemases as 6/6 at 4 h. There was no difference in results between the 4 h and 6 h time points. However, Blood-rsCDM could not differentiate OXA-181-producing strains. For eCIM, the characterization numbers for KPC-, OXA-181-, and MBLs-producing strains were 77/77, 2/2, and 57/59, respectively, but it failed to detect the coproduction of KPC and NDM isolates.ConclusionBlood-rsCDM accurately discriminates carbapenemase within 4 h and is capable of directly differentiating multi-enzyme (KPC and NDM) presence from positive blood culture broths. Therefore, Blood-rsCDM represents a rapid, simple, easy-to-read, and accurate tool that can be utilized in resource-limited settings.

Non-Invasive Detection of Nitrogen Deficiency in Cannabis sativa Using Hand-Held Raman Spectroscopy

Proper crop management requires rapid detection methods for abiotic and biotic stresses to ensure plant health and yield. Hemp (Cannabis sativa L.) is an emerging economically and environmentally sustainable crop capable of yielding high biomass. Nitrogen deficiency significantly reduces hemp plant growth, affecting photosynthetic capacity and ultimately decreasing yield. When symptoms of nitrogen deficiency are visible to humans, there is often already lost yield. A real-time, non-destructive detection method, such as Raman spectroscopy, is therefore critical to identify nitrogen deficiency in living hemp plant tissue for fast, precise crop remediation. A two-part experiment was conducted to investigate portable Raman spectroscopy as a viable hemp nitrogen deficiency detection method and to compare the technique’s predictive ability against a handheld SPAD (chlorophyll index) meter. Raman spectra and SPAD readings were used to train separate nitrogen deficiency discrimination models. Raman scans displayed characteristic spectral markers indicative of nitrogen deficiency corresponding to vibrational modes of carotenoids, essential pigments for photosynthesis. The Raman-based model consistently predicted nitrogen deficiency in hemp prior to the onset of visible stress symptoms across both experiments, while SPAD only differentiated nitrogen deficiency in the second experiment when the stress was more pronounced. Our findings add to the repertoire of plant stresses that hand-held Raman spectroscopy can detect by demonstrating the ability to provide assessments of nitrogen deficiency. This method can be implemented at the point of cultivation, allowing for timely interventions and efficient resource use.

Accurate identification of locally aneuploid cells by incorporating cytogenetic information in single cell data analysis

Single-cell RNA sequencing is a powerful tool to investigate the cellular makeup of tumor samples. However, due to the sparse data and the complex tumor microenvironment, it can be challenging to identify neoplastic cells that play important roles in tumor growth and disease progression. This is especially relevant for blood cancers, where neoplastic cells may be highly similar to normal cells. To address this challenge, we have developed partCNV and partCNVH, two methods for rapid and accurate detection of aneuploid cells with local copy number deletion or amplification. PartCNV uses an expectation-maximization (EM) algorithm with mixtures of Poisson distributions and incorporates cytogenetic information to guide the classification. PartCNVH further improves partCNV by integrating a hidden Markov model for feature selection. We have thoroughly evaluated the performance of partCNV and partCNVH through simulation studies and real data analysis using three scRNA-seq datasets from blood cancer patients. Our results show that partCNV and partCNVH have favorable accuracy and provide more interpretable results compared to existing methods. In the real data analysis, we have identified multiple biological processes involved in the oncogenesis of myelodysplastic syndromes and acute myeloid leukemia.

NanoMIP beacons with a co-operative binding mechanism for the all-in-one detection of methamphetamine aptamer complexes

Methamphetamine is a highly addictive stimulant with significant public health implications, necessitating the development of rapid, sensitive, and reliable detection methods. Traditional analytical techniques, though accurate, often involve complex sample preparation, expensive equipment, and lengthy analysis times. This study presents the design, synthesis, and application of nanoMIP beacons with a unique co-operative binding mechanism for the detection of methamphetamine. NanoMIP beacons selective for methamphetamine/aptamer complexes were synthesized using a solid-phase synthesis method that involved the bio-conjugation of an aptamer on the stationary phase and the incubation of methamphetamine to form a methamphetamine-aptamer complex. The resultant nanoMIP beacons were eluted off the affinity column and characterised using transmission electron microscopy. The co-operative binding mechanism, which relies on the structure-switching capability of the aptamer upon analyte binding was demonstrated through comparison of the fluorescence quenching signal of a scrambled sequence and nanoNIP controls. The fluorescence quenching assay was established using a fixed optimal concentration of aptamer and varying amounts of methamphetamine. The nanoMIP beacons showed enhanced the sensitivity (LOD = 23 ± 3.5 nM) and excellent selectivity, with a 40-fold increase in quenching for methamphetamine compared to other illicit drugs. The nanoMIP beacons demonstrated acceptable sample recoveries in both urine diluent and 50% human serum. This work provides a new strategy for the development of hybrid nanoMIP/aptamer-based sensors and provides a robust analytical tool for combating methamphetamine abuse.

Comparison of qPCR and chromogenic culture methods for rapid detection of group B streptococcus colonization in Vietnamese pregnant women

IntroductionNeonatal infections can rapidly become severe, with delays in treatment often proving fatal. Streptococcus agalactiae (Group B Streptococcus, GBS) is a common cause, typically transmitted from colonized pregnant women to neonates during childbirth. In Vietnam, routine prenatal care lacks standardized GBS screening protocols. This study aims to compare enhanced qPCR methods with the culture method, evaluate the diagnostic accuracy of these qPCR procedures, and assess the frequency of GBS infection in pregnant Vietnamese women during their final trimester. Materials and methodsPregnant women aged 35 weeks gestation or more were recruited. Rectovaginal swabs were collected and analyzed for GBS using chromogenic culture, a commercial real-time PCR kit, and in-house real-time PCR assays targeting the cfb and sip genes. Clinical diagnostic values were calculated, and GBS prevalence was determined. ResultsThe study included 259 pregnant women with a mean age of 30.2 ± 5.0 years. Of these, 96.6 % had gestational ages of 37 weeks or more at delivery. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the cfb-based and sip-based qPCR assays were 94.1/92.7, 99.0/99.5, 97.1/98.5, 97.8/97.3, and 97.6 %, respectively. The Kappa values were excellent (0.940 and 0.939), with results available in under 2 h. GBS prevalence was 24.7 % and 25.5 % by cfb-based and sip-based qPCR assays, aligning with the culture method (25.5 %). ConclusionsBoth direct real-time PCR assays demonstrated high accuracy and were comparable to chromogenic culture in diagnosing GBS. A significant prevalence of GBS colonization was found among Vietnamese pregnant women in their final trimester.

Research on Rapid Detection of Salmonella and Listeria monocytogenes Using Colloidal Gold Immunoassay and PCR Methods

This study used colloidal gold immunoassay strips and PCR (probe method) kit methods to detect Salmonella and Listeria monocytogenes. The accuracy and precision of the colloidal gold immunoassay strips and PCR (probe method) kit methods were validated using proficiency testing blind samples, and the consistency with the current national gold standard method was compared. The results of the two rapid detection methods were consistent with the results of the national standard method. This paper also analyzed the detection time and cost of the two rapid detection methods and the national standard method. The PCR (probe method) kit method takes less time for detection and can be used for high-throughput detection, with high sensitivity and low interference. It requires high equipment and personnel capabilities. The colloidal gold immunoassay strips method has the lowest detection cost, but its sensitivity is lower than that of the PCR method. It is easy to operate and requires low personnel and equipment requirements. Laboratories can choose the appropriate method based on their actual needs.

Development of a Rapid Detection Method for Ethylene Glycol and Glycolic Acid in Feline Samples: A Response to Increasing Antifreeze Poisoning Incidents in Korea

Recently, cases of antifreeze poisoning in companion animals, particularly cats, have surged in the Republic of Korea. Ethylene glycol (EG), the toxic primary component of antifreeze, is metabolized into glycolic acid (GA), leading to severe metabolic acidosis, acute kidney injury, and death. Traditional detection methods, although effective, are often time-consuming owing to complex sample preparation. This study involved a novel analytical method utilizing GC-MS for EG and LC-MS/MS for GA detection, which streamlined the detection process by eliminating the need for derivatization. The method was validated for accuracy and reliability, enabling the rapid and precise identification of EG and GA in biological samples. This study also included the successful application of this method in a case where initial exposure to antifreeze was not apparent, which highlighted the effectiveness of this method in diagnosing poisoning even in cases where clinical history is unclear. The development of this rapid diagnostic approach addresses the urgent need for the efficient detection of antifreeze poisoning, improving animal welfare and supporting forensic investigations.

Leaf Moisture Content Detection Method Based on UHF RFID and Hyperdimensional Computing

Leaf moisture content (LMC) directly affects the life activities of plants and becomes a key factor to evaluate the growth status of plants. To explore a low-cost, real-time, rapid, and accurate method for LMC detection, this paper employs Ultra-High-Frequency Radio-Frequency Identification (UHF RFID) sensor technology. By reading the tag information attached to the back of leaves, the parameters of the RSSI, phase, and reading distance of the tags are collected. In this paper, we propose an enhanced Multi-Feature Fusion algorithm based on Hyperdimensional Computing (HDC) called MFFHDC. In our proposed method, the real-valued features are encoded into hypervectors and then combined with Multi-Linear Discriminant Analysis (MLDA) for the feature fusion of different features. Finally, a retraining method based on Cosine Annealing with Warm Restarts (CAWR) is proposed to improve the model and further enhance its accuracy. Tests conducted in the experimental forest show that the proposed mechanism can effectively predict the LMC. The model’s Mean Absolute Error (MAE), Root Mean Square Error (RMSE), and Coefficient of Determination (R2) reached 0.0195, 0.0255, and 0.9131, respectively. Additionally, comparisons with other methods demonstrate that the presented system performs excellently in most aspects. As a lightweight model, this study shows great practical application value, particularly for the limited data volume and low hardware costs.

Enhancing Sensitivity and Selectivity: Current Trends in Electrochemical Immunosensors for Organophosphate Analysis.

This review examines recent advancements in electrochemical immunosensors for the detection of organophosphate pesticides, focusing on strategies to enhance sensitivity and selectivity. The widespread use of these pesticides has necessitated the development of rapid, accurate, and field-deployable detection methods. We discuss the fundamental principles of electrochemical immunosensors and explore innovative approaches to improve their performance. These include the utilization of nanomaterials such as metal nanoparticles, carbon nanotubes, and graphene for signal amplification; enzyme-based amplification strategies; and the design of three-dimensional electrode architectures. The integration of these sensors into microfluidic and lab-on-a-chip devices has enabled miniaturization and automation, while screen-printed and disposable electrodes have facilitated on-site testing. We analyze the challenges faced in real sample analysis, including matrix effects and the stability of biological recognition elements. Emerging trends such as the application of artificial intelligence for data interpretation and the development of aptamer-based sensors are highlighted. The review also considers the potential for commercialization and the hurdles that must be overcome for widespread adoption. Future research directions are identified, including the development of multi-analyte detection platforms and the integration of sensors with emerging technologies like the Internet of Things. This comprehensive overview provides insights into the current state of the field and outlines promising avenues for future development in organophosphate pesticide detection.

A Lab Prototype for Rapid Electrochemical Detection of Escherichia coli in Water Using Modified Screen-Printed Electrodes.

Recognizing the need for a hand-held device capable of quantitatively measuring the concentration of bacteria in water, this paper describes a label-free method for rapid detection of Escherichia coli (E. coli) in water via H2O2 decomposition using screen-printed electrodes (SPE) modified with nanostructured metal oxide layers. The study encompasses sensor preparation, bacteria culture, synthesis and characterization of nanostructures, and development of a readout circuitry for lab prototyping. During sensing measurements, the bacteria are first made to interact with H2O2 and subsequently, the H2O2 solution is exposed on the sensing surface. The electrochemical sensors are fabricated by modifying the working electrode of SPE with nanostructured metal oxide layers of MnO2 and TiO2 as these play a crucial role in the detection of E. coli in water. The sensing experiments of MnO2-modified SPE show a significant response to bacteria with a sensitivity of 0.82 mV.mL/log CFU and a limit of detection (LOD) of 1.8 CFU/mL, while the TiO2-modified SPE exhibits a linear response over a wide range of bacterial concentrations with a sensitivity of 1.12 mV·mL/log CFU and a limit of detection of 2.23 CFU/mL. Both sensors demonstrate a rapid response, stability, repeatability, and a recovery time of 70 ms. Additionally, selectivity with respect to other bacteria, wastewater components such as glucose, ammonium sulfate, and sodium carbonate, and testing with RO, DI, and tap water samples are conducted to evaluate the sensors' performance. A detailed sensing mechanism has been developed to comprehend the results, including chemical and biological reactions, metal oxide interfaces, morphology, and other surface studies of the sensing surface. A prototype comprising a sensor chip, an Arduino board, and other necessary circuit components is tested with various bacterial solutions. This enables its use for on-field rapid detection of bacteria in water using smaller volumes and a portable system.

Nanomaterials in bacterial detection: current trends and future outlook

Contamination by pathogenic bacteria represents a severe risk to public health and well-being. We outlined current approaches to detecting and sensing harmful bacteria by integrating recognition elements with nanomaterials (NMs) in this study. Nanomaterials have emerged as a transformative technology for bacterial detection due to their unique physicochemical properties, including high surface area, quantum effects, and enhanced reactivity. This review highlights the current trends in the application of various nanomaterials, such as gold nanoparticles, carbon nanotubes, and quantum dots, in the detection of bacterial pathogens. These materials enable the development of selective, and rapid detection methods through mechanisms like surface plasmon resonance, electrochemical sensing, and fluorescence. Furthermore, integrating nanomaterials with microfluidic devices and biosensors is discussed, showcasing advancements in point-of-care diagnostics. Challenges such as stability, reproducibility, and potential toxicity of nanomaterials are addressed, alongside regulatory considerations. The future outlook emphasizes the potential of emerging nanomaterials, such as graphene and metal-organic frameworks, to revolutionize bacterial detection. This review aims to enhance the scalability, cost-effectiveness, and environmental sustainability of these technologies, paving the way for widespread clinical and environmental applications.

Preparation and Application of Polyclonal Antibodies for the Rapid Detection of Actinidia Chlorotic Ringspot-Associated Virus

Actinidia chlorotic ringspot-associated virus (AcCRaV, Emaravirus actinidiae) is prevalent in Chinese kiwifruit, leading to substantial yield reduction. The intricate nature of symptoms presents diagnostic challenges, underscoring the necessity for a rapid and accurate detection method that facilitates effective control. In this investigation, AcCRaV isolates from key kiwi-producing regions in Sichuan province were collected and analyzed, with representative strains chosen as experimental materials. Primers targeting the nucleoprotein gene of AcCRaV were designed, and their codon usage was optimized to enhance performance. Various serological methods utilizing polyclonal antibodies were developed, including ELISA, dot immunobinding assay, and AcCRaV-specific gold immunochromatographic bands (AcCRaV-GICS). Field samples exhibited high specificity and sensitivity when tested using these methods. Furthermore, the results obtained from a large number of field samples are consistent with those derived from RT-PCR analysis, further validating the applicability of our approach. A detection method capable of handling a large volume of field samples infected with AcCRaV is currently lacking; thus, our system construction provides an important reference for addressing this gap.

Application of Multivariate Data Analysis Methods for Rapid Detection and Quantification of Adulterants in Lavender Essential Oil Using Infrared Spectroscopy

Application of Multivariate Data Analysis Methods for Rapid Detection and Quantification of Adulterants in Lavender Essential Oil Using Infrared Spectroscopy