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一起非典型肉毒毒素中毒事件实验室检测分析

Laboratory detection and analysis of an atypical botulism outbreak

  • 摘要:
    目的  对一起疑似肉毒毒素中毒事件进行实验室检测,为预防和快速鉴别肉毒毒素中毒提供实验依据。
    方法 按照国家标准GB4789.12—2016、GB 15193.2—2014《食品安全国家标准 食品毒理学实验室操作规范》和肉毒毒素诊断血清说明书进行样品处理和检测,采用小鼠生物试验法、诊断血清进行肉毒毒素检测和确证定型试验。
    结果 将114只SPF级雌性KM小鼠,分组后分别进行检出试验和确证分型试验,采用腹腔注射方式染毒。检出试验结果显示,除对照组外各样品受试组小鼠均呈现典型的肉毒毒素中毒症状,腌制生鱼–胰酶组、胃内容物组和胃内容物–胰酶组小鼠1~36 h内全部死亡;腌制生鱼组和血液组小鼠未见死亡。确证分型试验结果显示,腌制生鱼–胰酶液分别加A、B、C、D、E、F和A~F混合型诊断血清小鼠均出现死亡,而使用灭菌生理盐水(NS)稀释的1:1、1:2和1:3稀释液再作用于A、B、C、D、E、F和A~F混合型诊断血清,小鼠均未出现死亡,但各毒素对照组小鼠均出现死亡,各毒素灭活对照组均无死亡;胃内容物–胰酶液使用NS1:1稀释后分别加A、B、C、D、E、F和A~F混合型诊断血清小鼠均无死亡,但毒素对照组小鼠出现死亡,毒素灭活对照组无死亡。根据肉毒毒素诊断血清说明书,判定中毒不是由典型的A、B、C、D、E、F血清型肉毒毒素引起,初步判断腌制生鱼和胃内容物中含有其他异型或异种肉毒毒素。
    结论 此次食物中毒事件是由非A、B、C、D、E、F型肉毒毒素引起。

     

    Abstract:
    Objective To conduct laboratory detection on a suspected botulism incident and provide experimental evidence for the prevention and rapid identification of botulism.
    Methods Sample processing and testing were carried out according to the national standards GB4789.12–2016, GB 15193.2–2014 "National Food Safety Standard - Laboratory Operation Standards for Food Toxicology," and the instructions for botulinum toxin diagnostic serum. Mouse bioassay and diagnostic serum were used for botulinum toxin detection and confirmatory typing tests.
    Results A total of 114 SPF-grade female KM mice were used and divided into groups for detection and confirmatory typing tests. The mice were intoxicated via intraperitoneal injection. The detection test results showed that, except for the control group, animals in all sample groups exhibited typical botulism symptoms. All animals in the marinated raw fish-trypsin group, gastric contents group, and gastric contents-trypsin group died within 1–36 h. No deaths were observed in the marinated raw fish group or the blood group. The confirmatory typing test results showed that all animals in the marinated raw fish-trypsin group injected with type A, B, C, D, E, F, and A–F mixed diagnostic sera died. However, when the marinated raw fish-trypsin solution was diluted with sterile normal saline (NS) at ratios of 1:1, 1:2, and 1:3 and then reacted with type A, B, C, D, E, F, and A–F mixed diagnostic sera, no deaths occurred in the animals. Deaths were observed in all toxin control groups, but no deaths occurred in the toxin inactivation control groups. When the gastric contents-trypsin solution was diluted with NS at a 1:1 ratio and then reacted with type A, B, C, D, E, F, and A–F mixed diagnostic sera, no animal deaths occurred. However, deaths were observed in the toxin control group, while no deaths occurred in the toxin inactivation control group. According to the instructions for the botulinum toxin diagnostic serum, the poisoning was not caused by typical type A, B, C, D, E, or F botulinum toxin. It was preliminarily determined that the marinated raw fish and gastric contents contained other atypical or non-typical botulinum toxins.
    Conclusions This food poisoning incident was caused by non-A, -B, -C, -D, -E, or -F botulinum toxin.

     

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