Article(id=1321170640502108899, tenantId=1146029695717560320, journalId=1319373219989389464, issueId=1321170520196895605, articleNumber=null, orderNo=null, doi=10.11847/zgggws1145169, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1692374400000, receivedDateStr=2023-08-19, revisedDate=1727539200000, revisedDateStr=2024-09-29, acceptedDate=1733241600000, acceptedDateStr=2024-12-04, onlineDate=1792921226858, onlineDateStr=2026-10-25, pubDate=1741536000000, pubDateStr=2025-03-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792921226858, onlineIssueDateStr=2026-10-25, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792921226858, creator=13701087609, updateTime=1792921226858, updator=13701087609, issue=Issue{id=1321170520196895605, tenantId=1146029695717560320, journalId=1319373219989389464, year='2025', volume='41', issue='3', pageStart='257', pageEnd='384', issueExtLink='null', onlineDate='null', pubDate='1741536000000', pubDateStr='2025-03-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792921198175, creator='13701087609', updateTime=1792921198175, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=376, endPage=379, ext={EN=ArticleExt(id=1321170641311609572, articleId=1321170640502108899, tenantId=1146029695717560320, journalId=1319373219989389464, language=EN, title=Laboratory detection and analysis of an atypical botulism outbreak, columnId=1320427686514770171, journalTitle=Chinese Journal of Public Health, columnName=Survey and Report, runingTitle=null, highlight=null, articleAbstract=
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.

, authors=Huiyan QIN, Hui YANG, Jiehong ZHANG, Qingbo LI, authorsList=Huiyan QIN, Hui YANG, Jiehong ZHANG, Qingbo LI, authorCompany=null, correspAuthors=Jiehong ZHANG, authorNote=null, correspAuthorsNote=
ZHANG Jiehong, E-mail:
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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。

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覃辉艳(1981—),主任技师,硕士,研究方向:食品及保健食品毒理和功能检测

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张洁宏,E-mail:
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Test results

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组别小鼠
数/只
出现中毒
症状
死亡
腌制生鱼组330
腌制生鱼–胰酶组333
胃内容物组333
胃内容物–胰酶组333
血液1组330
血液2组330
明胶磷酸盐缓冲液对照组300
明胶磷酸盐缓冲液–胰酶对照组300
), ArticleFig(id=1321170987996004481, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1321170640502108899, language=CN, label=表1, caption=

检出试验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
组别小鼠
数/只
出现中毒
症状
死亡
腌制生鱼组330
腌制生鱼–胰酶组333
胃内容物组333
胃内容物–胰酶组333
血液1组330
血液2组330
明胶磷酸盐缓冲液对照组300
明胶磷酸盐缓冲液–胰酶对照组300
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一起非典型肉毒毒素中毒事件实验室检测分析
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覃辉艳 , 杨慧 , 张洁宏 *, * , 李庆波
中国公共卫生 | 调查报告与分析 2025,41(3): 376-379
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中国公共卫生 |调查报告与分析 2025 , 41 (3) : 376 -379
一起非典型肉毒毒素中毒事件实验室检测分析
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覃辉艳, 杨慧, 张洁宏*, * , 李庆波
作者信息
  • 广西壮族自治区疾病预防控制中心卫生毒理所,南宁 530028
通讯作者:
张洁宏,E-mail:
作者简介:

覃辉艳(1981—),主任技师,硕士,研究方向:食品及保健食品毒理和功能检测

Laboratory detection and analysis of an atypical botulism outbreak
Huiyan QIN, Hui YANG, Jiehong ZHANG , Qingbo LI
Affiliations
  • Guangxi Autonomous Regional Center for Disease Control and Prevention, Nanning 530028, China
  • Corresponding Author:
    ZHANG Jiehong, E-mail:
出版时间: 2025-03-10 doi: 10.11847/zgggws1145169
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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。

肉毒毒素  /  腌制生鱼  /  食物中毒  /  非典型  /  实验室检测
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.

botulinum toxin  /  marinated raw fish  /  food poisoning  /  atypical  /  laboratory detection
覃辉艳, 杨慧, 张洁宏, 李庆波. 一起非典型肉毒毒素中毒事件实验室检测分析. 中国公共卫生, 2025 , 41 (3) : 376 -379 . DOI: 10.11847/zgggws1145169
Huiyan QIN, Hui YANG, Jiehong ZHANG, Qingbo LI. Laboratory detection and analysis of an atypical botulism outbreak[J]. Chinese Journal of Public Health, 2025 , 41 (3) : 376 -379 . DOI: 10.11847/zgggws1145169
肉毒毒素中毒是由肉毒毒素引起以神经系统症状为主要表现的一种严重中毒反应,可由摄入被肉毒毒素污染的食品引起。肉毒毒素是由肉毒梭菌,有时还有丁酸梭菌和巴氏梭菌产生的一种神经毒素,其毒性极强,静脉注射1~3 ng/kg体重剂量、口服1 μg/kg体重剂量即可致人死亡[1]。根据抗原性的不同,肉毒毒素可分为A、B、C、D、E、F、G 7个血清型,2014年,Barash J R等[2]从一例肉毒毒素中毒的患儿粪便中分离出一种称为H血清型的新型肉毒毒素。肉毒毒素毒作用机制主要是通过抑制神经末梢乙酰胆碱的释放,导致肌肉麻痹和神经功能障碍,其中呼吸肌麻痹是其致死的主要原因[3]。
天然发生的食源性肉毒中毒主要是由于腌制的豆制品、腌肉等食品加工生产不当引起的,候荣丹[4]报道6例患者因食用自制豆腐乳而发生A型肉毒毒素中毒,常文广等[5]报道5例患者因食用自制臭豆腐和自腌的豆制品造成B型肉毒毒素中毒。此外,食用加工不当的腌制牛肉可致B、E型肉毒毒素中毒[6]。在婴儿肉毒毒素中毒中除可见B型[7]外,还可见由丁酸梭菌引起的E型[8]和巴氏梭菌引起的F型[9]肉毒毒素中毒。目前,国内外食源性肉毒毒素中毒由A、B、E型引起的报道较多,F型偶见,但其他型的报道较少。本研究对一起2024年2月19日广西某医院报告的疑似食用自制腌制生鱼引起的肉毒毒素中毒事件进行实验室检测,以确认是否为腌制食品细菌肉毒毒素中毒及毒素分型,结果报告如下。
2024年2月19日广西某医院报告了2例疑似食物中毒患者,患者主要以呕吐、腹胀、头晕乏力、视物模糊、呼吸困难等症状为主。经调查核实,2月15日14时患者家庭7人聚餐,其中5人食用了自制的腌制生鱼,从2月16日05时起,陆续有3人出现上述症状,其中重症1例,无死亡病例。2月20日采样送检了同批次自制的腌制生鱼1罐(未曾食用),2名患者血液样品各2管,胃内容物1管。
SPF级雌性KM小鼠114只,体质量15~20 g,由广西医科大学医学实验动物中心提供[SCXK(桂)2020-0003],实验动物质量合格证号为2024026。动物饲料由广西医科大学医学实验动物中心提供[SCXK(桂)2020–0003] ,动物实验室为屏障系统[SYXK(桂)2021–0002]。实验动物室温度22~25 ℃,相对湿度55%~70%。动物试验方案经过广西壮族自治区疾病预防控制中心实验动物管理与伦理委员会审查批准(审查批号:20240001)。
数显三用恒温水浴箱(金坛亿通电子公司);生物安全柜(德国Thermo公司);Sigma 4-16K高速冷冻离心机(德国SIGMA公司)。肉毒毒素诊断血清(兰州生物制品研究所有限责任公司);生理盐水(normal saline,NS)。
参照GB 4789.12—2016《食品安全国家标准 食品微生物学检验 肉毒梭菌及肉毒毒素检验》[10] 、GB 15193.2—2014《食品安全国家标准 食品毒理学实验室操作规范》[11] 和肉毒毒素诊断血清说明书[12]方法进行样品处理和检测。
无菌操作称取腌制生鱼样品25 g,放入无菌乳钵,研杵研碎,加入50 mL明胶磷酸盐缓冲液,混匀,浸泡30 min,取样品匀液放入离心管,3 000 r/min离心20 min,收集上清液,取部分上清液加10%胰酶(v:v,9:1)处理,胰酶活力1:250;胃内容物离心后取上清液,部分上清液加10%胰酶(v:v,9:1)处理;血液样品离心后取血清;供试验用。另取明胶磷酸盐缓冲液加10%胰酶(v:v,9:1)处理,供试验对照使用。
将雌性KM小鼠24只分为腌制生鱼组、腌制生鱼–胰酶组、胃内容物组、胃内容物–胰酶组、血液1组、血液2组、明胶磷酸盐缓冲液对照组和明胶磷酸盐缓冲液–胰酶对照组共8组,每组3只。分别取腌制生鱼处理液和腌制生鱼–胰酶处理液、胃内容物上清液和胃内容物–胰酶处理液、病例血清(1号和2号)、明胶磷酸盐缓冲液、明胶磷酸盐缓冲液–胰酶处理液给予相应组别小鼠腹腔注射,每只0.5 mL,观察和记录小鼠48 h内的中毒表现。
根据检出试验结果,将雌性KM小鼠90只分为5个大组,分别为腌制生鱼–胰酶液(原液)组、腌制生鱼–胰酶与NS1:1稀释液组、腌制生鱼–胰酶与NS1:2稀释液组、腌制生鱼–胰酶与NS1:3稀释液组和胃内容物–胰酶与NS1:1稀释液组,每个大组18只小鼠。每个大组再分别分为9个小组,分别为A、B、C、D、E、F、A~F混合、毒素对照和毒素灭活对照组,每组2只小鼠。将腌制生鱼–胰酶液使用灭菌NS分别稀释成1:1、1:2、1:3稀释液,将胃内容物–胰酶液使用灭菌NS稀释成1:1稀释液,将腌制生鱼–胰酶液和上述稀释液分别与A、B、C、D、E、F血清和A~F 6型混合血清等量混合,37℃孵育45 min,分别给予相应组别小鼠腹腔注射,每只0.5 mL,观察和记录96 h内小鼠发病与死亡情况。同时各毒素对照组分别腹腔注射各受试液与灭菌NS等量混合液,各毒素灭活对照组分别腹腔注射煮沸20 min灭活的各受试液与灭菌NS等量混合液[12]。此次试验步骤主要参照肉毒毒素诊断血清说明书,与国家标准GB 4789.12—2016[10]在孵育时间、灭活时间和稀释液上稍有差别,本实验室之前一直使用此品牌试剂盒,根据之前的试剂盒验证试验结果和同类检验的经验,本试验采用了试剂盒标示的方法进行了本次检测。
腌制生鱼组和腌制生鱼–胰酶组、胃内容物组和胃内容物–胰酶组腹腔注射小鼠5 min后陆续可见受试小鼠出现腹部凹陷宛如蜂腰、竖毛、呼吸急促、活动减少等中毒症状,腌制生鱼–胰酶组小鼠在给受试物后1~2 h内死亡,胃内容物组和胃内容物–胰酶组小鼠在给受试物后1~36 h内死亡。腌制生鱼组小鼠24 h后陆续恢复正常,无死亡。血液组腹腔注射小鼠20 min后,受试小鼠陆续出现竖毛、活动减少、呼吸急促中毒症状,症状较轻,6 h后恢复正常,未见动物死亡。明胶磷酸盐缓冲液对照组和明胶磷酸盐缓冲液–胰酶对照组受试小鼠均未见中毒症状和死亡。
腹腔注射各受试液10 min后,除各毒素灭活对照组外各组受试小鼠陆续出现竖毛、腰腹部凹陷、呼吸急促、活动减少等中毒症状,各稀释液组症状较轻。腌制生鱼–胰酶液(原液)A、B、D、E、F、A~F混合型组和毒素对照组全部小鼠和C组半数小鼠2 h后陆续死亡,毒素灭活对照组未见小鼠死亡;腌制生鱼–胰酶与NS 1:1、1:2和1:3稀释液A、B、C、D、E、F、A~F混合型组和毒素灭活对照组均未见小鼠死亡,而腌制生鱼–胰酶与NS1:1稀释液毒素对照组全部小鼠和腌制生鱼–胰酶与NS 1:2和1:3稀释液毒素对照组半数小鼠20 h后陆续死亡;胃内容物–胰酶与NS 1:1稀释液A、B、C、D、E、F、A~F混合型组和毒素灭活对照组均未见小鼠死亡,而毒素对照组全部小鼠20 h后陆续死亡。
目前,肉毒毒素的检测方法有小鼠生物试验法、ELISA、免疫色谱法、免疫PCR法和细胞学检测法等多种方法[13],小鼠生物试验法仍是公认的肉毒毒素检测“金标准”,该方法能评估活性毒素的整体功能,且灵敏度高,可达10 pg/mL[14]。本研究采用小鼠生物试验法对患者自制的腌制生鱼、胃内容物、血液样品进行肉毒毒素检测,从毒素的来源(腌制生鱼)、消化(胃内容物)、吸收(血液)多个层面探讨此次食物中毒。
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,腌制生鱼加胰酶处理后可致小鼠死亡,未加胰酶处理未见小鼠死亡,表明该毒素本身毒性较低,可被胰酶激活,致毒性增强。胃内容物不论是否加胰酶处理均可致小鼠死亡,表明该毒素已在体内被激活,故可引起患者中毒症状。两份血液样品虽可致小鼠呈现肉毒毒素中毒症状,但未致小鼠死亡,表明血液中的毒素含量较低,原因可能为该毒素在体内被激活后不能100%吸收进入血液循环,且血液样品为患者中毒5天后采集,血液中的毒素已经过机体一定的代谢分解和排泄,故含量较低。此外,从上述检出试验结果可以看出,患者中毒症状轻重除与进食中毒食物数量有关外,也与机体胰酶的活性和吸收代谢排泄相关。
进一步的确证分型试验采用肉毒毒素诊断血清,其原理为抗原与抗体特异性结合使得特异型肉毒毒素在小鼠体内呈无毒性症状。结果显示:(1)各毒素组受试小鼠均出现死亡,而各毒素灭活组均无死亡,除原液混合型组小鼠出现死亡外,其余各稀释液混合型组均无死亡,表明该毒素可被热灭活,浓度不高时可被肉毒毒素混合血清中和,表明腌制生鱼样品和胃内容物中含有肉毒毒素。(2)腌制生鱼–胰酶液(原液)A、B、C、D、E、F和混合型组小鼠均出现死亡,而1:1、1:2和1:3稀释液A、B、C、D、E、F和混合型组小鼠均未出现死亡,表明各型血清均可中和样品中一定量的毒素,但如样品毒素量较高,则各型血清和混合血清均无法中和。根据肉毒毒素诊断血清说明书,单一型血清每mL可中和相应型毒素至少10 000 LD50,混合型血清每mL可中和各型毒素至少10 000 LD50,表明各型血清和混合型血清可中和其相应型的毒素能力很强,达到半数致死量的10 000倍。样品1:3稀释(4倍稀释)后,只能引起小鼠半数致死,表明受试样品原液的毒素含量远远低于10 000倍的半数致死量,但各型血清和混合型血清均未能中和样品原液中的毒素,均引起小鼠死亡,表明样品中的毒素不是A、B、C、D、E和F型;另根据诊断血清说明书,单一型血清每mL可中和异型或异种毒素不高于5 LD50,混合型血清每mL可中和异种毒素不高于5 LD50,表明各型血清和混合血清均可中和一定量的异型或异种毒素(不高于半数致死量的5倍)。试验中当样品进行稀释后,各型血清和混合血清均可中和样品稀释液中的毒素,未引起小鼠死亡,表明各稀释液可能含有不高于5 LD50的异型或异种毒素;如使用样品原液则各型血清和混合血清均无法中和样本原液中的毒素而引起小鼠死亡,表明样品原液可能含有的异型或异种毒素高于半数致死量的5倍。因此,初步判断腌制生鱼样品中含有其他异型或异种肉毒毒素。同理,胃内容物–胰酶与NS1:1稀释液毒素组受试小鼠全部死亡,而A、B、C、D、E、F和混合型组小鼠均无死亡,表明各型血清均可中和样品中的毒素,根据肉毒毒素诊断血清说明书,初步判断胃内容物中含有其他异型或异种肉毒毒素。
本次试验结果显示,此次中毒事件不是由典型的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。目前关于其他型肉毒毒素的研究报道较少,Sonnabend O等[15]报道从5例食物中毒骤死病人分离到G型肉毒杆菌及其毒素,G型肉毒毒素亦具有不耐热,可被胰酶激活的特性。Terilli R R等[16]亦报道G型生物的毒性相对较低,在培养中仅产生少量毒素,这一特性使其很难被识别,需要胰酶激活才能被成功检测到。并且产毒菌株在几次培养传代后会丧失产生G型肉毒毒素的能力,较难检测到[17]。Barash J R等[2]从一例婴儿肉毒毒素中毒患者粪便中分离出的新型H型肉毒毒素可被热灭活,未见是否可被胰酶激活的报道。本研究检测到的肉毒毒素与G型一样有不耐热和可被胰酶激活的特性,但因目前市面上没有G型和H型肉毒毒素的诊断血清,本研究无法进一步检测探讨该肉毒毒素是否为G型或H型,或是其他异型肉毒毒素。新型毒素的鉴定有赖于菌株分离和型别鉴定,因此在突发事件早期处置过程中应尽可能多地保存中毒样品和污染样品,并严格按照病原微生物的保存要求做好保存,以便在日后的毒素检测过程中如果发现为疑似新型毒素,可以进一步做好菌株分离和型别鉴定,为新型毒素确认提供更明确依据的同时,也为下一步开发新型别的毒素鉴定试剂盒提供实物条件。
本研究采用小鼠生物试验法对此次疑似肉毒毒素食物中毒进行检测,患者胃内容物和同批次自制腌制生鱼均检出非A、B、C、D、E、F型肉毒毒素,血液样品毒素含量低,无法检出。此次样品均为患者进食自制的腌制生鱼5天后所采集,在中毒中后期采集剩余食物样品和患者胃内容物等胃肠道样品,比血液样品更具检测价值。对于所进食的有毒食物,如无法采集吃剩样品,同批次制作样品亦具有检测价值。另小鼠生物试验法虽是肉毒毒素检测“金标准”,但在本研究中也发现了该方法一定的局限性,即需要的样品量较大;阳性结果能在24 h内得出,但需要96 h确证阴性样品或低浓度样品;受诊断血清限制,无法判定更多的相应型别的肉毒毒素。

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doi: 10.11847/zgggws1145169
  • 接收时间:2023-08-19
  • 首发时间:2026-10-25
  • 出版时间:2025-03-10
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  • 收稿日期:2023-08-19
  • 修回日期:2024-09-29
  • 录用日期:2024-12-04
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    广西壮族自治区疾病预防控制中心卫生毒理所,南宁 530028

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2种不同金属材料的力学参数
科
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属数
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genus
种数
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species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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