High-resolution mass spectrometry and enzyme-linked immunosorbent assay for detection of residual botulinum concentrations in a mouse model of botulism

IF 1.4 4区医学 Q3 MEDICINE, LEGAL

Legal Medicine Pub Date : 2026-02-01 Epub Date: 2025-11-19 DOI:10.1016/j.legalmed.2025.102758

Dongqing Li , Junyan Liang , Ying Chen , Xianglei Wu

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引用次数: 0

Abstract

Background

Botulinum toxin type A is widely used to block acetylcholine release in the treatment of chronic sialorrhea, muscle spasticity, and dystonia. We aim to develop a user-friendly method for detecting cases of medical botulinum toxin poisoning.

Methods

The mice poisoning model was established by injecting or gavage with Botulax®, and the poisoning dose and symptoms were observed. The residual levels of toxin in poisoned mice were detected by high-resolution mass spectrometry and sandwich ELISA, respectively.

Results

Two hours after poisoning, no residual botulinum toxin was found by mass spectrometry (MS) under our specific untargeted workflow and sample preparation conditions, but ELISA detected residual toxin in various tissues of mice. Among them, the muscle tissue had the highest level. There is no noticeable difference in the levels of toxin residues in the same organs of mice, regardless of the route of poisoning. The sandwich ELISA method is user-friendly for detecting medical botulinum toxin poisoning. The presence of toxin residues can be detected in various tissues two hours after exposure, with muscle being the optimal sampling tissue.

Conclusions

Our research indicates that under specific sample preparation, chromatographic separation, and untargeted detection conditions, mass spectrometry may not be effective for detecting Botulinum toxin at concentrations below the ng/ml level. The study demonstrates that ELISA is a sensitive and practical alternative for early detection. Positive results can be detected within 2 h of poisoning, especially when taken from muscle tissue.

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高分辨率质谱法和酶联免疫吸附法检测肉毒杆菌中毒小鼠模型中的残留肉毒杆菌浓度

背景A型肉毒杆菌毒素被广泛用于阻断乙酰胆碱的释放，用于治疗慢性唾液分泌、肌肉痉挛和肌张力障碍。我们的目的是开发一种用户友好的方法来检测病例的医用肉毒杆菌毒素中毒。方法采用肉毒毒素®注射或灌胃法建立小鼠中毒模型，观察中毒剂量和中毒症状。分别采用高分辨率质谱法和夹心ELISA法检测中毒小鼠体内毒素残留水平。结果在特定的非靶向工作流程和样品制备条件下，中毒2 h后，质谱法（MS）未检测到肉毒毒素残留，而ELISA法在小鼠各组织中检测到肉毒毒素残留。其中，肌肉组织中含量最高。无论中毒途径如何，在小鼠的同一器官中，毒素残留水平没有明显差异。夹心ELISA法是检测医用肉毒毒素中毒的简便方法。暴露后两小时可在各种组织中检测到毒素残留物的存在，肌肉是最佳取样组织。结论在特定的样品制备、色谱分离和非靶向检测条件下，质谱法可能无法检测ng/ml以下浓度的肉毒杆菌毒素。该研究表明，ELISA是一种灵敏、实用的早期检测方法。中毒后2小时内可检测出阳性结果，特别是从肌肉组织中提取时。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Legal Medicine Nursing-Issues, Ethics and Legal Aspects

CiteScore

2.80

自引率

6.70%

发文量

119

审稿时长

7.9 weeks

期刊介绍： Legal Medicine provides an international forum for the publication of original articles, reviews and correspondence on subjects that cover practical and theoretical areas of interest relating to the wide range of legal medicine. Subjects covered include forensic pathology, toxicology, odontology, anthropology, criminalistics, immunochemistry, hemogenetics and forensic aspects of biological science with emphasis on DNA analysis and molecular biology. Submissions dealing with medicolegal problems such as malpractice, insurance, child abuse or ethics in medical practice are also acceptable.