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Keywords = colloidal gold immunoassay strips

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14 pages, 2042 KB  
Article
Boosting Sensitivity, Stability, and Speed: A Polydopamine-Engineered Silver Nanoparticle Lateral Flow Immunoassay for Aflatoxin B1 in Maize
by Xinge Mo, Shuhong Zhang, Zixuan He, Xiaoyang Li, Xiangmin Li, Yonghua Xiong and Hu Jiang
Toxins 2026, 18(3), 129; https://doi.org/10.3390/toxins18030129 - 3 Mar 2026
Viewed by 763
Abstract
Conventional colorimetric lateral flow immunoassays (LFIAs) often suffer from insufficient sensitivity for detecting trace low-molecular-weight contaminants like mycotoxins. The development of colorimetric probes with a high molar extinction coefficient is therefore critical for enhancing detection performance. Although silver nanoparticles (AgNPs) exhibit an extremely [...] Read more.
Conventional colorimetric lateral flow immunoassays (LFIAs) often suffer from insufficient sensitivity for detecting trace low-molecular-weight contaminants like mycotoxins. The development of colorimetric probes with a high molar extinction coefficient is therefore critical for enhancing detection performance. Although silver nanoparticles (AgNPs) exhibit an extremely high molar extinction coefficient, their practical application in LFIA is hindered by inherent chemical instability and suboptimal visual contrast. To address these limitations, we have engineered robust and high-performance polydopamine-functionalized AgNPs (Ag@PDA NPs) as advanced LFIA signal probes, which were successfully used for detecting aflatoxin B1 (AFB1) in maize. The multifunctional PDA nanoshell effectively shields the Ag core from oxidation and other destabilizing factors, ensuring superior long-term stability and significantly enhancing colorimetric contrast. Moreover, it improves the colloidal hydrophilicity, enabling faster and more uniform migration kinetics along the test strip. Leveraging these engineered properties, the developed assay achieved a limit of detection (LOD) of 0.23 ng mL−1 for AFB1 in buffer, representing a remarkable 2.17-fold sensitivity enhancement over conventional colloidal gold-based LFIAs. Validation in spiked maize samples confirmed high reliability, with recoveries ranging from 95.70% to 119.28% and precision (inter-/intra-assay CVs) below 13.03%. Full article
(This article belongs to the Section Mycotoxins)
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16 pages, 942 KB  
Article
Association Study on Risk Factors for Major Infectious Diseases in Dogs and Cats in Shenzhen, China
by Yao Peng, Runchang Lin, Wanxing Xie, Rongjie Huang, Shunping Cai, Yinyi Liang, Qida Lin, Gen Li, Xiaofeng Guo, Bowen Lin and Jun Luo
Animals 2026, 16(1), 49; https://doi.org/10.3390/ani16010049 - 24 Dec 2025
Viewed by 1903
Abstract
This study investigated the prevalence of 11 common pathogens in dogs and cats in Shenzhen, China, from January 2022 to March 2024, aiming to enhance the understanding of their epidemiological characteristics for improved disease control strategies. Diagnostic testing for the target pathogens was [...] Read more.
This study investigated the prevalence of 11 common pathogens in dogs and cats in Shenzhen, China, from January 2022 to March 2024, aiming to enhance the understanding of their epidemiological characteristics for improved disease control strategies. Diagnostic testing for the target pathogens was performed using polymerase chain reaction (PCR), colloidal gold test strips, or fluorescence immunoassay. Statistical analysis revealed that among 13,134 cats, Feline Panleukopenia Virus (FPV) showed the highest prevalence (35.83%), followed by Feline Calicivirus (FCV, 26.20%), Feline Infectious Peritonitis Virus (FIPV, 22.00%), and Feline Herpesvirus (FHV, 15.76%). Among 3626 dogs, Canine Parvovirus (CPV) and Canine Distemper Virus (CDV) were predominant, showing a prevalence of 54.55% and 42.83%, respectively. Risk factor analysis showed that most infections occurred in unvaccinated animals and young individuals (<1 year old), with higher incidences in winter and spring. Logistic regression indicated that sex, age, and season were significantly associated with FPV, FHV, and FIPV infections, while age and season were associated with FCV, CPV, and CDV infections (sex showed no association). This study contributes to the epidemiological knowledge of common infectious diseases in dogs and cats, providing a theoretical basis for disease prevention in dogs and cats. Full article
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11 pages, 1773 KB  
Brief Report
Development of a Nanogold-Based Lateral Flow Immunoassay for Point-of-Care Detection of SARS-CoV-2 Nucleocapsid Proteins and Antibodies
by Wei-Jie Tsai, Yeh Chen, Jye-Lin Hsu, Hsiao-Chuan Lin, Po-Ren Hsueh and Cheng-Wen Lin
COVID 2025, 5(9), 158; https://doi.org/10.3390/covid5090158 - 18 Sep 2025
Viewed by 2656
Abstract
The ongoing COVID-19 pandemic has underscored the urgent need for rapid, sensitive, and versatile diagnostic tools. In this study, we developed a nanogold-based lateral flow immunoassay (LFIA) capable of detecting both SARS-CoV-2 nucleocapsid (N) protein antigens and anti-N IgG antibodies at the point [...] Read more.
The ongoing COVID-19 pandemic has underscored the urgent need for rapid, sensitive, and versatile diagnostic tools. In this study, we developed a nanogold-based lateral flow immunoassay (LFIA) capable of detecting both SARS-CoV-2 nucleocapsid (N) protein antigens and anti-N IgG antibodies at the point of care. Following optimization of colloidal gold nanoparticle size, pH, and protein conjugation parameters, LFIA strips were assembled in two formats: a competitive assay for antigen detection and a sandwich assay for antibody detection. In the competitive format, gold nanoparticles (AuNPs)-conjugated N protein were used to detect varying concentrations of free N protein. The test line signal inversely correlated with antigen concentration, confirming the assay’s specificity and effectiveness. For antibody detection, the sandwich LFIA format employed immobilized anti-human IgG to capture anti-N antibodies in serum samples from COVID-19 patients. Strong test line signals were observed in samples collected ≥11 days post-symptom onset, indicating a time-dependent increase in IgG detectability. These results demonstrate that the AuNP-based LFIA platform provides a flexible, rapid, and low-cost diagnostic solution, suitable for both early antigen detection and serological monitoring during SARS-CoV-2 infection and recovery. Full article
(This article belongs to the Section COVID Clinical Manifestations and Management)
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26 pages, 57341 KB  
Article
AI-Powered Embedded System for Rapid Detection of Veterinary Antibiotic Residues in Food-Producing Animals
by Ximing Li, Lanqi Chen, Qianchao Wang, Mengting Zhou, Jingheng Long, Xi Chen, Jiangsan Zhao, Junjun Yu and Yubin Guo
Antibiotics 2025, 14(9), 917; https://doi.org/10.3390/antibiotics14090917 - 11 Sep 2025
Cited by 1 | Viewed by 1823
Abstract
Background: Veterinary antibiotics are widely used in food-producing animals, raising public health concerns due to drug residues and the risk of antimicrobial resistance. Rapid and reliable detection systems are critical to ensure food safety and regulatory compliance. Colloidal gold immunoassay (CGIA)-based antigen–antibody test [...] Read more.
Background: Veterinary antibiotics are widely used in food-producing animals, raising public health concerns due to drug residues and the risk of antimicrobial resistance. Rapid and reliable detection systems are critical to ensure food safety and regulatory compliance. Colloidal gold immunoassay (CGIA)-based antigen–antibody test cards are widely used in food safety for the rapid screening of veterinary antibiotic residues. However, manual interpretation of test cards remains inefficient and inconsistent. Methods: To address this, we propose a complete AI-based detection system for veterinary antibiotic residues. The system is built on the Rockchip RK3568 platform and integrates a five-megapixel OV5640 autofocus USB camera (60° field of view) with a COB LED strip (6000 K, rated 5 W/m). It enables high-throughput, automated interpretation of colloidal gold test cards and can generate structured detection reports for regulatory documentation and quality control. The core challenge lies in achieving accurate and fast inference on resource-constrained embedded devices, where traditional detection networks often struggle to balance model size and performance. To this end, we propose VetStar, a lightweight detection algorithm specifically optimized for this task. VetStar integrates StarBlock, a shallow feature extractor, and Depthwise Separable-Reparameterization Detection Head (DR-head), a compact, partially decoupled detection head that accelerates inference while preserving accuracy. Results: Despite its compact size, with only 0.04 M parameters and 0.3 GFLOPs, VetStar maintains strong performance after distillation with the Bridging Cross-task Protocol Inconsistency Knowledge Distillation (BCKD) method. For our custom Veterinary Drug Residue Rapid Test Card (VDR-RTC) dataset, it achieves an mAP50 of 97.4 and anmAP50-95of 89.5. When deployed on the RK3568 device, it delivers results in just 5.4 s—substantially faster than comparable models. Conclusions: These results highlight the system’s strong potential for high-throughput, cost-effective, and rapid veterinary antibiotic residue screening, supporting food safety surveillance efforts. Full article
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11 pages, 3073 KB  
Article
Au Nanoshell-Based Lateral Flow Immunoassay for Colorimetric and Photothermal Dual-Mode Detection of Interleukin-6
by Congying Wen, Yue Dou, Yao Liu, Xuan Jiang, Xiaomei Tu and Ruiqiao Zhang
Molecules 2024, 29(15), 3683; https://doi.org/10.3390/molecules29153683 - 3 Aug 2024
Cited by 12 | Viewed by 3460
Abstract
Interleukin-6 (IL-6) detection and monitoring are of great significance for evaluating the progression of many diseases and their therapeutic efficacy. Lateral flow immunoassay (LFIA) is one of the most promising point-of-care testing (POCT) methods, yet suffers from low sensitivity and poor quantitative ability, [...] Read more.
Interleukin-6 (IL-6) detection and monitoring are of great significance for evaluating the progression of many diseases and their therapeutic efficacy. Lateral flow immunoassay (LFIA) is one of the most promising point-of-care testing (POCT) methods, yet suffers from low sensitivity and poor quantitative ability, which greatly limits its application in IL-6 detection. Hence, in this work, we integrated Aushell nanoparticles (NPs) as new LFIA reporters and achieved the colorimetric and photothermal dual-mode detection of IL-6. Aushell NPs were conveniently prepared using a galvanic exchange process. By controlling the shell thickness, their localized surface plasmon resonance (LSPR) peak was easily tuned to near-infrared (NIR) range, which matched well with the NIR irradiation light. Thus, the Aushell NPs were endowed with good photothermal effect. Aushell NPs were then modified with IL-6 detection antibody to construct Aushell probes. In the LFIA detection, the Aushell probes were combined with IL-6, which were further captured by the capture IL-6 antibody on the test line of the strip, forming a colored band. By observation with naked eyes, the colorimetric qualitative detection of IL-6 was achieved with limit of 5 ng/mL. By measuring the temperature rise of the test line with a portable infrared thermal camera, the photothermal quantitative detection of IL-6 was performed from 1~1000 ng/mL. The photothermal detection limit reached 0.3 ng/mL, which was reduced by nearly 20 times compared with naked-eye detection. Therefore, this Aushell-based LFIA efficiently improved the sensitivity and quantitative ability of commercial colloidal gold LFIA. Furthermore, this method showed good specificity, and kept the advantages of convenience, speed, cost-effectiveness, and portability. Therefore, this Aushell-based LFIA exhibits practical application potential in IL-6 POCT detection. Full article
(This article belongs to the Special Issue Functional Nanomaterials for Biosensors and Biomedicine Application)
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12 pages, 3870 KB  
Article
Rapid and Sensitive Detection of Influenza B Virus Employing Nanocomposite Spheres Based on Ag-Doped ZnIn2S4 Quantum Dots
by Jia-Xuan Hu, Li-Bang Zhu, Sheng-Tong Wu and Shou-Nian Ding
Chemosensors 2024, 12(4), 68; https://doi.org/10.3390/chemosensors12040068 - 19 Apr 2024
Cited by 6 | Viewed by 3241
Abstract
Lateral flow immunoassay (LFIA) technology serves a significant role as a simple and rapid biosensor in the detection of influenza viruses. The focus of this study is the development of a rapid and convenient screening method for influenza B virus (IBV) proteins using [...] Read more.
Lateral flow immunoassay (LFIA) technology serves a significant role as a simple and rapid biosensor in the detection of influenza viruses. The focus of this study is the development of a rapid and convenient screening method for influenza B virus (IBV) proteins using a fluorescence lateral flow biosensor based on Ag-doped ZnIn2S4 quantum dots (Ag: ZIS QDs) as signal reporters. These Ag: ZIS QDs-emitting orange fluorescence are loaded onto dendritic mesoporous silica nanoparticles (DMSNs) and are further coated with a layer of silica shell to form a core–shell structured composite nanomaterial (SiO2 @ Ag: ZIS QDs @ DMSNs). The orange fluorescence effectively eliminates the interference of blue background fluorescence, significantly enhancing the detection sensitivity. This technology demonstrates outstanding performance in the immediate detection of IBV, with a minimum detection limit of 1 ng/mL, compared to the traditional colloidal gold strip with a detection limit of 6 ng/mL. Furthermore, both intra-assay and inter-assay coefficients of variation (CV) are less than 9%. This method holds promise for wide application in early diagnosis, epidemiological investigation, and epidemic surveillance of IBV. Full article
(This article belongs to the Special Issue Rapid Point-of-Care Testing Technology and Application)
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13 pages, 2418 KB  
Article
In Situ Synthesis of Highly Fluorescent, Phosphorus-Doping Carbon-Dot-Functionalized, Dendritic Silica Nanoparticles Applied for Multi-Component Lateral Flow Immunoassay
by Jia-Xuan Hu and Shou-Nian Ding
Sensors 2024, 24(1), 19; https://doi.org/10.3390/s24010019 - 19 Dec 2023
Cited by 7 | Viewed by 3487
Abstract
The sensitivity of fluorescent lateral flow immunoassay (LFIA) test strips is compromised by the low fluorescence intensity of the signaling molecules. In this study, we synthesized novel phosphorus-doped carbon-dot-based dendritic mesoporous silica nanoparticles (DMSNs-BCDs) with a quantum yield as high as 93.7% to [...] Read more.
The sensitivity of fluorescent lateral flow immunoassay (LFIA) test strips is compromised by the low fluorescence intensity of the signaling molecules. In this study, we synthesized novel phosphorus-doped carbon-dot-based dendritic mesoporous silica nanoparticles (DMSNs-BCDs) with a quantum yield as high as 93.7% to break this bottleneck. Meanwhile, the in situ growth method increased the loading capacity of carbon dots on dendritic mesoporous silica, effectively enhancing the fluorescence intensity of the composite nanospheres. Applied DMSNs-BCDs in LFIA can not only semi-quantitatively detect a single component in a short time frame (procalcitonin (PCT), within 15 min) but also detect the dual components with a low limit of detection (LOD) (carbohydrate antigen 199 (CA199) LOD: 1 U/mL; alpha-fetoprotein (AFP) LOD: 0.01 ng/mL). And the LOD of PCT detection (0.01 ng/mL) is lower by 1.7 orders of magnitude compared to conventional colloidal gold strips. For CA199, the LOD is reduced by a factor of four compared to LFIA using gold nanoparticles as substrates, and for AFP, the LOD is lowered by two orders of magnitude compared to colloidal gold LFIA. Furthermore, the coefficients of variation (CV) for intra-assay and inter-assay measurements are both less than 11%. Full article
(This article belongs to the Special Issue Fluorescence Sensors for Biological and Medical Applications)
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12 pages, 4535 KB  
Article
Lateral Flow Immunoassay Based on Quantum-Dot Nanobeads for Detection of Chloramphenicol in Aquatic Products
by Qian Han, Ling Fan, Xiuying Liu, Yiwei Tang, Pingping Wang, Zaixi Shu, Wei Zhang and Lijie Zhu
Molecules 2023, 28(22), 7496; https://doi.org/10.3390/molecules28227496 - 9 Nov 2023
Cited by 18 | Viewed by 3905
Abstract
Quantum dot nanobeads (QBs) were used as signal source to develop competitive lateral flow immunoassay (LFIA) for the detection of chloramphenicol (CAP). The quantitative detection of CAP was achieved by calculating the total color difference (∆E) values of the test line (T line) [...] Read more.
Quantum dot nanobeads (QBs) were used as signal source to develop competitive lateral flow immunoassay (LFIA) for the detection of chloramphenicol (CAP). The quantitative detection of CAP was achieved by calculating the total color difference (∆E) values of the test line (T line) using the images of test strips. QB-based LFIA (QBs-LFIA) allowed the effective dynamic linear detection of CAP in the range of 0.1–1.5 ng/mL. The limit of detection (LOD) was 3.0 ng/mL, which was 50 and 667 times lower than those achieved for two different brands of colloidal gold kits. The recoveries of CAP during real-sample detection were 82.82–104.91% at spiked levels of 0.1, 0.7, and 1.5 ng/mL. These results indicate that the developed QBs-LFIA facilitates the sensitive detection of CAP. Full article
(This article belongs to the Section Materials Chemistry)
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11 pages, 1591 KB  
Article
Rapid Colloidal Gold Immunoassay for Pharmacokinetic Evaluation of Vancomycin in the Cerebrospinal Fluid and Plasma of Beagle Dogs
by Yechang Guo, Shaofeng Wang, Peiyue Li, Pan Zhang and Wei Wang
Sensors 2023, 23(21), 8978; https://doi.org/10.3390/s23218978 - 5 Nov 2023
Cited by 1 | Viewed by 3168
Abstract
Vancomycin (VAN), a glycopeptide antibiotic, is the preferred therapeutic agent for treating Gram-positive bacteria. Rapid and precise quantification of VAN levels in cerebrospinal fluid (CSF) and plasma is crucial for optimized drug administration, particularly among elderly patients. Herein, we introduce a novel clinical [...] Read more.
Vancomycin (VAN), a glycopeptide antibiotic, is the preferred therapeutic agent for treating Gram-positive bacteria. Rapid and precise quantification of VAN levels in cerebrospinal fluid (CSF) and plasma is crucial for optimized drug administration, particularly among elderly patients. Herein, we introduce a novel clinical test strip utilizing colloidal gold competitive immunoassay technology for the expedient detection of VAN. This test strip enables the detection of VAN concentrations in clinical samples such as plasma within 10 min and has a limit of detection of 10.3 ng/mL, with an inhibitory concentration 50% (IC50) value of 44.5 ng/mL. Furthermore, we used the test strip for pharmacokinetic analysis of VAN in the CSF and plasma of beagle dogs. Our results provide valuable insights into the fluctuations of the drug concentration in the CSF and plasma over a 24 h period after a single intravenous dose of 12 mg/kg. The test strip results were compared with the results obtained via liquid chromatography–mass spectrometry methods, and the measured VAN concentrations in the CSF and plasma via both of the methods showed excellent agreement. Full article
(This article belongs to the Section Biomedical Sensors)
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12 pages, 2733 KB  
Article
Magnetic SERS Strip Based on 4-mercaptophenylboronic Acid-Modified Fe3O4@Au for Active Capture and Simultaneous Detection of Respiratory Bacteria
by Jingfei Li, Jin Chen, Yuwei Dai, Zhenzhen Liu, Junnan Zhao, Shuchen Liu and Rui Xiao
Biosensors 2023, 13(2), 210; https://doi.org/10.3390/bios13020210 - 31 Jan 2023
Cited by 21 | Viewed by 5514 | Correction
Abstract
The rapid diagnosis and detection of respiratory bacteria at the early stage can effectively control the epidemic spread and bacterial infection. Here, we designed a rapid, ultrasensitive, and quantitative lateral flow immunoassay (LFA) strip for simultaneous detection of respiratory bacteria S. aureus and [...] Read more.
The rapid diagnosis and detection of respiratory bacteria at the early stage can effectively control the epidemic spread and bacterial infection. Here, we designed a rapid, ultrasensitive, and quantitative lateral flow immunoassay (LFA) strip for simultaneous detection of respiratory bacteria S. aureus and S. pneumoniae. In this assay, the surface enhanced Raman scattering (SERS) tags were designed through combining magnetite Raman enhancement nanoparticle Fe3O4@Au/DTNB and recognition element 4-mercaptophenylboronic acid (4-MPBA). Further, 4-MPBA could capture multiple bacteria in a complex environmental solution. Based on the strategies, Fe3O4@Au/DTNB-mediated magnetic enrichment and 4-MPBA-mediated universal capture capabilities improved the detection sensitivity, the limits of detection for S. aureus and S. pneumoniae were as low as 12 and 11 CFU mL−1, respectively, which were more sensitive than those of colloidal gold method. The Fe3O4@Au/DTNB/Au/4-MPBA-LFA also exhibited good reproducibility, excellent specificity, and high recovery rates in sputum samples, indicating its potential application in the detection of respiratory bacteria samples. Full article
(This article belongs to the Section Biosensors and Healthcare)
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15 pages, 3164 KB  
Article
Preparation of Monoclonal Antibody against Deoxynivalenol and Development of Immunoassays
by Hoyda Elsir Mokhtar, Aidi Xu, Yang Xu, Mohamed Hassan Fadlalla and Shihua Wang
Toxins 2022, 14(8), 533; https://doi.org/10.3390/toxins14080533 - 3 Aug 2022
Cited by 14 | Viewed by 3370
Abstract
Fusarium toxins are the largest group of mycotoxins, which contain more than 140 known secondary metabolites of fungi. Deoxynivalenol (DON) is one of the most important compounds of this class due to its high toxicity and its potential to harm mankind and animals [...] Read more.
Fusarium toxins are the largest group of mycotoxins, which contain more than 140 known secondary metabolites of fungi. Deoxynivalenol (DON) is one of the most important compounds of this class due to its high toxicity and its potential to harm mankind and animals and a widespread contaminant of agricultural commodities, such as wheat, corn, barley, oats, bread, and biscuits. Herein, a hybridoma cell 8G2 secreting mAb against DON was produced by fusing the splenocytes with a tumor cell line Sp2/0. The obtained mAb had a high affinity (2.39 × 109 L/mol) to DON. An indirect competitive Enzyme-Linked Immunosorbent Assay (ic-ELISA) showed that the linear range for DON detection was 3.125–25 μg/mL, and the minimum inhibitory concentration (IC50) was 18.125 μg/mL with a limit of detection (LOD) of 7.875 μg/mL. A colloidal gold nanoparticle (AuNP) with 20 nm in diameter was synthesized for on-site detection of DON within 10 min with vLOD of 20 μg/mL. To improve the limit of detection, the gold nanoflower (AuNF) with a larger size (75 nm) was used to develop the AuNF-based strip with vLOD of 6.67 μg/mL. Compared to the vLOD of a convectional AuNP-based strip, the AuNF-based strip was three times lower. Herein, three immunoassay methods (ic-ELISA and AuNP/AuNF-based strips) were successfully developed, and these methods could be applied for the DON detection in agricultural products. Full article
(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China (2nd Edition))
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12 pages, 6483 KB  
Article
Development and Application of a Visual Duck Meat Detection Strategy for Molecular Diagnosis of Duck-Derived Components
by Xiaoyun Chen, Huiru Yu, Yi Ji, Wei Wei, Cheng Peng, Xiaofu Wang, Xiaoli Xu, Meihao Sun and Junfeng Xu
Foods 2022, 11(13), 1895; https://doi.org/10.3390/foods11131895 - 26 Jun 2022
Cited by 12 | Viewed by 3394
Abstract
To make meat adulteration detection systems faster, simpler and more efficient, we established a duck-derived meat rapid detection Recombinase Polymerase Amplification (dRPA) method by using interleukin 2 (IL-2) from nuclear genomic DNA as the target gene to design specific primers. We tested the [...] Read more.
To make meat adulteration detection systems faster, simpler and more efficient, we established a duck-derived meat rapid detection Recombinase Polymerase Amplification (dRPA) method by using interleukin 2 (IL-2) from nuclear genomic DNA as the target gene to design specific primers. We tested the dRPA detection system by comparing its sensitivity and specificity using real-time fluorescent PCR technology. By adjusting the ratio of reagents, this method shortens the time of DNA extraction and visualizes results in combination with colloidal gold immunoassay strips. Our results demonstrate that this dRPA method could specifically detect duck-derived components with a sensitivity of up to 23 copies/μL and the accuracy of the results is consistent with real-time fluorescent PCR. Additionally, dRPA can detect at least 1% of the duck meat content by mixing beef and mutton with duck meat in different proportions, which was verified by spot-check market samples. These results can be visualized with colloidal gold immunoassay strips with the same accuracy as real-time fluorescent RPA. dRPA can complete detection within 30 min, which shortens existing detection time and quickly visualizes the detection results on-site. This lays the groundwork for future large-scale standardized duck origin detection. Full article
(This article belongs to the Special Issue PCR in Food Science: Current Technology and Applications)
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17 pages, 1140 KB  
Protocol
The Strategic Alliance between Clinical and Molecular Science in the War against SARS-CoV-2, with the Rapid-Diagnostics Test as an Indispensable Weapon for Front Line Doctors
by Antonio Vittorino Gaddi, Fabio Capello, Leonardo Aluigi, Pier Luigi Antignani, Annapaola Callegaro, Gavino Casu, Enrico Cipolla, Maurizio Cipolla, Lucio Cosco, Federico Culzoni, Francesco Dentali, Maria Elexpuru-Zabaleta, Tamara Y. Forbes-Hernandez, Claudia Fragiacomo, Francesca Giampieri, Agostino Gnasso, Raffaele Mancini, Maria Grazia Modena, Michele Nichelatti, Angelo Virgilio Paradiso, Pasquale Ortasi, Maria Teresa Savo, Flavio Tangianu, Sergio Tempesta, Tommaso Diego Voci and Maurizio Battinoadd Show full author list remove Hide full author list
Int. J. Mol. Sci. 2020, 21(12), 4446; https://doi.org/10.3390/ijms21124446 - 22 Jun 2020
Cited by 8 | Viewed by 5677
Abstract
Our work concerns the actual problem of spread of SARS- CoV-2 outbreak which requires fast and correct as possible answer. In current scenario, the need of rapid answer put away the imperative of proper methodology. We focus on the serogical immunoassay for diagnosis [...] Read more.
Our work concerns the actual problem of spread of SARS- CoV-2 outbreak which requires fast and correct as possible answer. In current scenario, the need of rapid answer put away the imperative of proper methodology. We focus on the serogical immunoassay for diagnosis of Covid-19 as an important weapon not only for diagnostic purpose, but also for epidemiologic one. The right equilibrium between high speed, low cost and accuracy is obtained with easy-to-use decentralized point-of-care test as the colloidal gold-based immunochromatographic strip assay which detects IgM and IgG antibodies directed against SARS-CoV-2. As our aim is to evaluate the efficacy of Covid-19 rapid tests and of serological assays in real-life settings, we designed a research protocol aimed to establish how to use correctly these diagnostics, taking into account the different possible clinical and epidemiological scenarios. Full article
(This article belongs to the Section Molecular Biology)
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11 pages, 2124 KB  
Article
Preparation of Monoclonal Antibody for Brevetoxin 1 and Development of Ic-ELISA and Colloidal Gold Strip to Detect Brevetoxin 1
by Sumei Ling, Shiwei Xiao, Chengjie Xie, Rongzhi Wang, Linmao Zeng, Ke Wang, Danping Zhang, Xiulan Li and Shihua Wang
Toxins 2018, 10(2), 75; https://doi.org/10.3390/toxins10020075 - 8 Feb 2018
Cited by 26 | Viewed by 6368
Abstract
Brevetoxin-1 (BTX-1), a marine toxin mostly produced by the dinoflagellatae Karenia brevis, has caused the death of marine organisms and has had numerous toxicological effects on human health. Hence, it is very necessary to develop a rapid, economical, and reliable immunoassay method [...] Read more.
Brevetoxin-1 (BTX-1), a marine toxin mostly produced by the dinoflagellatae Karenia brevis, has caused the death of marine organisms and has had numerous toxicological effects on human health. Hence, it is very necessary to develop a rapid, economical, and reliable immunoassay method for BTX-1 detection. In this study, two kinds of complete antigen were synthesized using the succinic anhydride and isobutyl chloroformate two-step methods. Conjugate BTX-1-OVA was used as an antigen for mice immunization, and BTX-1-BSA for measuring the titer of the produced antibodies. A hybridoma cell line 6C6 stably secreting monoclonal antibody (mAb) against BTX-1 was obtained by fusing SP2/0 myeloma cells with the spleen cells from the immunized mouse. The hybridoma 6C6 was injected into the abdomen of BALB/c mice to obtain ascites, and the anti-BTX-1 mAb was harvested from ascites by precipitation with caprylic acid/ammonium sulfate (CA-AS). The anti-BTX-1 mAb was identified as an IgG1 subtype, and the cross-reactivity results showed that anti-BTX-1 mAb was highly specific to BTX-1 with the affinity of 1.06 × 108 L/mol. The indirect competitive ELISA results indicated that the linear range for BTX-1 detection was 14–263 ng/mL with IC50 of 60 ng/mL, and a detection limit of 14 ng/mL. The average recovery rate from the spiked samples was 88 ± 2% in intra-assay and 89 ± 2% in inter-assay. The limit of detection (LOD) using the colloidal gold strip was 200 ng/mL with high specificity. Therefore, the anti-BTX-1 mAb can be used to detect BTX-1 in shellfish and other related samples. Full article
(This article belongs to the Section Marine and Freshwater Toxins)
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10 pages, 1351 KB  
Article
Rapid Detection of Escherichia coli O157 and Shiga Toxins by Lateral Flow Immunoassays
by Jinliang Wang, Robab Katani, Lingling Li, Narasimha Hegde, Elisabeth L. Roberts, Vivek Kapur and Chitrita DebRoy
Toxins 2016, 8(4), 92; https://doi.org/10.3390/toxins8040092 - 25 Mar 2016
Cited by 38 | Viewed by 12135
Abstract
Shiga toxin-producing Escherichia coli O157:H7 (STEC) cause food-borne illness that may be fatal. STEC strains enumerate two types of potent Shiga toxins (Stx1 and Stx2) that are responsible for causing diseases. It is important to detect the E. coli O157 and Shiga toxins [...] Read more.
Shiga toxin-producing Escherichia coli O157:H7 (STEC) cause food-borne illness that may be fatal. STEC strains enumerate two types of potent Shiga toxins (Stx1 and Stx2) that are responsible for causing diseases. It is important to detect the E. coli O157 and Shiga toxins in food to prevent outbreak of diseases. We describe the development of two multi-analyte antibody-based lateral flow immunoassays (LFIA); one for the detection of Stx1 and Stx2 and one for the detection of E. coli O157 that may be used simultaneously to detect pathogenic E. coli O157:H7. The LFIA strips were developed by conjugating nano colloidal gold particles with monoclonal antibodies against Stx1 and Stx2 and anti-lipid A antibodies to capture Shiga toxins and O157 antigen, respectively. Our results indicate that the LFIA for Stx is highly specific and detected Stx1 and Stx2 within three hours of induction of STEC with ciprofloxacin at 37 °C. The limit of detection for E. coli O157 LFIA was found to be 105 CFU/mL in ground beef spiked with the pathogen. The LFIAs are rapid, accurate and easy to use and do not require sophisticated equipment or trained personnel. Following the assay, colored bands on the membrane develop for end-point detection. The LFIAs may be used for screening STEC in food and the environment. Full article
(This article belongs to the Collection Rapid Detection of Bacterial Toxins)
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