孔雀石绿的磁性免疫层析快速检测方法研究

开发了一种适用于现场快速检测孔雀石绿(MG)的免疫层析试纸条,在超顺磁性纳米微球上偶联MG单克隆抗体作为检测探针,分别将孔雀石绿完全抗原(MG-B SA)和羊抗鼠IgG喷涂于NC膜的T线和C线.结果 发现,T线最佳喷涂量为0.25 mg/mL,抗体最佳偶联量为20 μg,构建的试纸条可在25 min内实现养殖用水及鱼肉...     

Monoclonal Antibodies Application in Lateral Flow Immunochromatographic Assays for Drugs of Abuse Detection

Lateral flow assays (lateral flow immunoassays and nucleic acid lateral flow assays) have experienced a great boom in a wide variety of early diagnostic and screening applications. As opposed to conventional examinations (High Performance Liquid Chromatography, Polymerase Chain Reaction, Gas chromatography-Mass Spectrometry, etc.), they obtain the results of a sample’s analysis within a short period. In resource-limited areas, these tests must be simple, reliable, and inexpensive. In this review, we outline the production process of antibodies against drugs of abuse (such as heroin, amphetamine, benzodiazepines, cannabis, etc.), used in lateral flow immunoassays as revelation or detection molecules, with a focus on the components, the principles, the formats, and the mechanisms of reaction of these assays. Further, we report the monoclonal antibody advantages over the polyclonal ones used against drugs of abuse. The perspective on aptamer use for lateral flow assay development was also discussed as a possible alternative to antibodies in view of improving the limit of detection, sensitivity, and specificity of lateral flow assays.     

不同类型探针对免疫层析方法的分析灵敏度影响研究

为研究不同类型探针对免疫层析方法的分析灵敏度影响,该研究以17β-雌二醇(17β-E2)为检测对象,采用磁性纳米材料标记7μg/mL的17β-E2单克隆抗体(检测抗体)制备了传统探针,采用磁性纳米材料分别标记5μg/mL的17β-E2单克隆抗体和5μg/mL的羊抗鼠IgG多克隆抗体(第二抗体)制备了配对探针,采用磁性纳米材料标记14μg/mL的羊抗鼠IgG多克隆抗体制备了间接探针。然后,基于这3种不同类型的探针,分别建立了17β-E2的免疫层析快速检测方法,并对3种17β-E2免疫层析方法的检测灵敏度进行比较。结果显示,基于传统、配对和间接探针的免疫层析方法对17β-E2的检出限分别为1、0.5、0.2 ng/mL,其中基于间接探针的免疫层析方法灵敏度最高,与基于传统探针和配对探针的免疫层析方法相比,检出限分别降低了5倍和2.5倍。该研究刷新了传统免疫层析方法中探针必须是信号材料标记检测抗体的认知,可为其他食品安全危害因子的免疫层析快速检测研究提供有价值的参考。     

重金属铬离子胶体金免疫层析快速检测法的建立

建立了水样中铬离子(Cr3+)检测的免疫试纸条检测方法。将自制的抗Cr3+-EDTA单克隆抗体标记于金颗粒后,包被于玻璃纤维上,将Cr3+-EDTA-BSA抗原和羊抗鼠IgG二抗分别结合于硝酸纤维膜上作为检测线(T线)和质控线(C线),组装试纸条。该试纸条可检测经亚硫酸氢钠还原为Cr3+的Cr6+,检出限为50μg/L;与10种重金属进行交叉实验验证,除与1 000μg/L的Fe3+有微弱交叉外,与其他重金属均无交叉反应;比较试纸条与仪器法ICP检测加标水样,两种方法的结果呈正相关。该试纸条可在3～5 min内肉眼观察结果,适用于现场检测,满足水样或土壤中重金属铬残留的监测要求。     

莱克多巴胺荧光胶乳颗粒免疫层析检测法的建立

基于免疫竞争层析法原理,建立了莱克多巴胺(RAC)的荧光胶乳颗粒免疫层析快速检测技术,用于猪肉组织中莱克多巴胺残留的检测.应用自制的抗RAC单抗,标记己胶乳荧光颗粒,将标记好的复合物喷涂于结合垫上.羊抗鼠IgG与人工合成的RAC - BSA检测抗原包被在NC膜表面,分别作为质控线(C线)与检测线(T线).检测过程中RA...     

基于不同纳米材料的侧流免疫层析技术在真菌毒素检测中的应用

真菌毒素毒性强,具有很强的致癌性和致畸性,对人体的危害巨大.近年来,其快速检测技术成为研究的热点,在“批量检测”的趋势下,侧流免疫层析技术以具有操作简便,经济合理,灵敏快速等优点,广泛应用于食品、饲料、农产品中真菌毒素的检测.本文对侧流免疫层析检测技术的研究概况及不同纳米材料的侧流免疫层析技术在真菌毒素检测中的应用和发展前景进行综述,以期为侧流免疫层析技术在真菌毒素检测的未来发展及应用提供参考.     

Gold nanoparticle-based immunochromatographic assay for the detection of 7-aminoclonazepam in urine

An analytical system of immunochromatographic assay based on gold nanoparticles was developed for the detection of 7-aminoclonazepam (7-ACLZ) in human urine. The qualitative assay was based on the competitive immunoassay using anti-7-ACLZ polyclonal antibody (PcAb) and a detector reagent that contains colloidal gold particles coated with anti-7-ACLZ PcAb. Nitrocellulose membrane was separately immobilised with goat anti-rabbit IgG (control line) and 7-ACLZ-OVA conjugate (test line). The sensitivity of the strip was tested for detecting 7-ACLZ spiked in urine and each specimen was independently measured by liquid chromatography tandem mass spectrometry. Good correlation was showed by the recovery results. The limit of detection for the strip test in urine was 100 ng mL^?1. The assay can be applied to the rapid detection of 7-ACLZ with the short testing time.     

截短新型冠状病毒N蛋白在荧光层析法上的应用

基于截短新型冠状病毒(Severe Acute Respiratory Syndrome Coronavirus 2,SARS-CoV-2)N蛋白和全长N蛋白,采用荧光免疫层析技术,建立了一种高灵敏度快速评价新型冠状病毒肺炎患者体内SARS-CoV-2抗体水平的方法。以荧光微球为标记物制备试纸条,进行阴性和阳性检测并选出较优抗原配对组合。初步评价了其精密度与稳定性,并与已上市的检测方法进行对比。根据阴性和阳性检测结果优选截短N蛋白标记,全长N蛋白包被组合,所建立的方法可以在 15 min内完成SARS-CoV-2抗体水平的检测,能够有效区分阴阳性,批内精密度CV为3.48%~10.05%,批间精密度CV为4.77%~11.73%。试纸条在37 ℃条件下能够稳定保存21 d,与胶体金检测法一致性较好,总符合率为96.13%,阳性符合率为90.63%,阴性符合率为97.56%。该研究基于截短N蛋白初步建立的荧光免疫层析检测方法能有效评价患者体内的SARS-CoV-2抗体水平,具有良好的精密度和稳定性,便于在基层医疗单位推广,有较好的应用前景。     

Development and evaluation of a rapid lateral flow immunochromatographic strip assay for screening 19-nortestosterone

The lateral flow strip test for 19-nortestosterone is one kind of immunochromatographic assay. Nitrocellulose membrane was separately immobilized with goat anti-rabbit IgG (control line) and 19-NT-OVA conjugate (test line). Anti-19-NT polyclonal antibody labeled with colloidal gold particles acted as the detector reagent. The assay is qualitatively, not quantitatively, judged with positive or negative result. We tested the sensitivity of the strip using spiked swine urine, and each specimen was independently measured by LC/MS/MS. The sensitivity, measure by eye, was determined to be 200 ng/mL. The assay time was less than 15 min, and so suitable for on-site rapid test.