RESUMEN

Halosulfuron-methyl (HM) is widely used for the removal of noxious weeds in corn, sugarcane, wheat, rice, and tomato fields. Despite its high efficiency and low toxicity, drift to nontarget crops and leaching of its metabolites to groundwater pose potential risks. Considering the instability of HM, the pyrazole sulfonamide of HM was used to generate a hapten and antigen to raise a high-quality monoclonal antibody (Mab, designated 1A91H11) against HM. A direct competitive immunoassay (dcELISA) using Mab 1A91H11 achieved a half-maximal inhibitory concentration (IC50) of 1.5 × 10-3 mg/kg and a linear range of 0.7 × 10-3 mg/kg-10.7 × 10-3 mg/kg, which was 10 times more sensitive than a comparable indirect competitive ELISA (icELISA) and more simple to operate. A spiking recovery experiment performed in tomato and maize matrices with 0.01, 0.05, and 0.1 mg/kg HM had average recoveries within 78.9-87.9% and 103.0-107.4% and coefficients of variation from 1.1-6.8% and 2.7-6.4% in tomato and maize, respectively. In addition, a magnetic lateral flow immunoassay (MLFIA) was developed for quantitative detection of low concentrations of HM in paddy water. Compared with dcELISA, the MLFIA exhibited 3.3- to 50-fold higher sensitivity (IC50 0.21 × 10-3 mg/kg). The average recovery and RSD of the developed MLFIA ranged from 81.5 to 92.5% and 5.4 to 9.7%. The results of this study demonstrated that the developed dcELISA and MLFIA are suitable for rapid detection of HM residues in tomato and maize matrices and paddy water, respectively, with acceptable accuracy and precision.

RESUMEN

In this study, we present a novel and ultrasensitive magnetic lateral flow immunoassay (LFIA) tailored for the precise detection of zearalenone, a mycotoxin with significant implications for human and animal health. A versatile and straightforward method for creating non-covalent magnetic labels is proposed and comprehensively compared with a covalent immobilization strategy. We employ the magnetic particle quantification (MPQ) technique for precise detection of the labels and characterization of their functionality, including measuring the antibody sorption density on the particle surface. Through kinetic studies using the label-free spectral phase interferometry, the rate and equilibrium constants for the binding of monoclonal antibodies with free (not bound with carrier protein) zearalenone were determined to be kon = 3.42 × 105 M-1s-1, koff = 7.05 × 10-4 s-1, and KD = 2.06 × 10-9 M. The proposed MPQ-LFIA method exhibits detection limits of 2.3 pg/mL and 7.6 pg/mL when employing magnetic labels based on covalent immobilization and non-covalent sorption, with dynamic ranges of 5.5 and 5 orders, correspondingly. We have successfully demonstrated the effective determination of zearalenone in barley flour samples contaminated with Fusarium graminearum. The ease of use and effectiveness of developed test systems further enhances their value as practical tools for addressing mycotoxin contamination challenges.

Asunto(s)

Micotoxinas , Zearalenona , Animales , Humanos , Zearalenona/análisis , Cinética , Micotoxinas/análisis , Inmunoensayo/métodos , Contaminación de Alimentos/análisis , Fenómenos Magnéticos , Límite de Detección

RESUMEN

Colorectal cancer (CRC) is the third leading cause of cancer death and the fourth most common cancer in the world. Colonoscopy is the most sensitive test used for detection of CRC; however, their procedure is invasive and expensive for population mass screening. Currently, the fecal occult blood test has been widely used as a screening tool for CRC but displays low specificity. The lack of rapid and simple methods for mass screening makes the early diagnosis and therapy monitoring difficult. Extracellular vesicles (EVs) have emerged as a novel source of biomarkers due to their contents in proteins and miRNAs. Their detection would not require invasive techniques and could be considered as a liquid biopsy. Specifically, it has been demonstrated that the amount of CD147 expressed in circulating EVs is significant higher for CRC cell lines than for normal colon fibroblast cell lines. Moreover, CD147-containing EVs have been used as a biomarker to monitor response to therapy in patients with CRC. Therefore, this antigen could be used as a non-invasive biomarker for the detection and monitoring of CRC in combination with a Point-of-Care platform as, for example, Lateral Flow Immunoassays (LFIAs). Here, we propose the development of a quantitative lateral flow immunoassay test based on the use of magnetic nanoparticles as labels coupled to inductive sensor for the non-invasive detection of CRC by CD147-positive EVs. The results obtained for quantification of CD147 antigen embedded in EVs isolated from plasma sample have demonstrated that this device could be used as a Point-of-Care tool for CRC screening or therapy monitoring thanks to its rapid response and easy operation.

Asunto(s)

Neoplasias Colorrectales , Vesículas Extracelulares , Biomarcadores de Tumor , Neoplasias Colorrectales/diagnóstico , Detección Precoz del Cáncer , Humanos , Inmunoensayo , Fenómenos Magnéticos

RESUMEN

BACKGROUND: Dengue infection represents a global health issue of growing importance. Dengue non-structural protein 1 (NS1) plays a central role in the early detection of the disease. The most common method for NS1 detection is testing by lateral flow immunoassays (LFIAs) with varying sensitivity. In this study, we present a highly sensitive magneto-enzyme LFIA for prompt diagnosis of dengue. METHODS: We have demonstrated the development of a magneto-enzyme LFIA combining super-paramagnetic nanoparticles as labels and Biotin-Streptavidin signal amplification strategy to detect dengue NS1. Factors affecting the test performance including antibody pair, super-paramagnetic nanoparticle size, nitrocellulose membrane type, amounts of detection and capture antibodies, and amounts of Streptavidin-polyHRP were optimized. Analytical sensitivity and cross-reactivity were determined. Clinical performance of the novel assay was evaluated using a panel of 120 clinical sera. RESULTS: This newly developed assay could detect NS1 of all four serotypes of dengue virus (DENV). The limit of detection (LOD) was found to be as low as 0.25 ng ml-1 for DENV-1 and DENV-3, 0.1 ng ml-1 for DENV-2, and 1.0 ng ml-1 for DENV-4. The LOD for DENV-2 was a 50-fold improvement over the best values previously reported. There was an absence of cross-reactivity with Zika NS1, Hepatitis B virus, Hepatitis C virus, and Japanese encephalitis virus. The sensitivity and specificity of the novel assay were 100% when tested on clinical samples. CONCLUSIONS: We have successfully developed a magneto-enzyme LFIA, allowing rapid and highly sensitive detection of dengue NS1, which is essential for proper management of patients infected with DENV.