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基于荧光的莨菪碱检测免疫层析测试条

Fluorescence-based immunochromatographic test strip for the detection of hyoscyamine.

机构信息

State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People's Republic of China.

Wuxi No. 2 People's Hospital, Wuxi, 214002, Jiangsu, 214122, People's Republic of China.

出版信息

Analyst. 2022 Jan 17;147(2):293-302. doi: 10.1039/d1an01973b.

Abstract

Hyoscyamine (HSM), which acts as an antagonist of the acetylcholine muscarinic receptor and can induce a variety of distinct toxic syndromes in mammals (anti-cholinergic poisoning), is hazardous to human health. Therefore, it is urgent to develop a rapid, sensitive, and cost-effective method to determine HSM. A fluorescent microsphere based immunochromatographic assay was developed for this analyte and gold nanoparticles (AuNPs) were used as a comparison. A monoclonal antibody against HSM was prepared with a 50% inhibition concentration (IC) of 1.17 ng mL, with no cross-reactivity with five drugs. Under optimized conditions, the cut off limits using the fluorescence-labeled monoclonal antibody strips were 10 ng mL in 0.01 M PBS and 20 ng mL in pork, pig urine, and honey samples, and the assay could be completed within 10 min. In comparison with a AuNP immunochromatographic assay, the developed method offered a higher coupling rate and lower amounts of antibodies. This approach could be used for simple, sensitive and rapid screening, and is suitable for on-site screening applications.

摘要

莨菪碱(HSM)作为乙酰胆碱毒蕈碱受体的拮抗剂,可在哺乳动物中诱导多种不同的毒性综合征(抗胆碱能中毒),对人类健康有害。因此,迫切需要开发一种快速、灵敏、经济有效的方法来测定 HSM。为此,开发了一种基于荧光微球的免疫层析分析方法,并将纳米金(AuNPs)作为比较。制备了一种针对 HSM 的单克隆抗体,其 50%抑制浓度(IC)为 1.17ng mL,与五种药物无交叉反应。在优化条件下,使用荧光标记的单克隆抗体条的截止值在 0.01 M PBS 中为 10ng mL,在猪肉、猪尿和蜂蜜样品中为 20ng mL,检测时间可在 10min 内完成。与 AuNP 免疫层析分析相比,该方法具有更高的偶联率和更低的抗体用量。该方法可用于简单、灵敏、快速的筛选,适用于现场筛选应用。

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