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Article(id=1151881497364541909, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250226001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1740499200000, receivedDateStr=2025-02-26, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752559549895, onlineDateStr=2025-07-15, pubDate=1748102400000, pubDateStr=2025-05-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752559549895, onlineIssueDateStr=2025-07-15, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752559549895, creator=13701087609, updateTime=1752559549895, updator=13701087609, issue=Issue{id=1151881493552394994, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='10', pageStart='1', pageEnd='324', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752559548986, creator=13701087609, updateTime=1756202008453, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1167159075906265916, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1167159075906265917, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=186, endPage=191, ext={EN=ArticleExt(id=1151923900981064537, articleId=1151881497364541909, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Enzyme-linked immunosorbent assay for detecting soybean agglutinin associated with kidney bean poisoning, columnId=1151895322692776479, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Analysis and Monitoring of Toxic and Harmful Substances in Food, runingTitle=null, highlight=null, articleAbstract=

Objective To establish an enzyme-linked immunosorbent assay method for detecting soybean agglutinin in legume foods. Methods After washed the sample, added phosphate buffer solution and homogenize it thoroughly. After centrifugation and purification, collected the supernatant and determine it by enzyme-linked immunosorbent assay. Quantify it used an enzyme-linked immunosorbent assay reader. Result In the linear range of 0-400 μg/mL, the linear equation of soybean agglutinin obtained was Y=0.0021X+0.0528, r=0.9998. The detection limit of this method was 125 mg/kg, the recovery rate was between 99.17% and 101.20%, and the relative standard deviation was between 1.27%-2.24%. Conclusion This method is easy to operate, has good specificity, and the antigen concentration is directly proportional to the OD450 value. Applicable to the detection of soybean agglutinin poisoning incidents by grassroots disease control centers.

, correspAuthors=Ying-Qian ZHANG, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Feng CHEN, En-De ZHONG, Xiao-Tian MING, Ying-Qian ZHANG, Jing-Jing ZHANG), CN=ArticleExt(id=1151923909067681896, articleId=1151881497364541909, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=酶联免疫吸附法检测芸豆中毒相关的大豆凝集素, columnId=1151923892102197877, journalTitle=食品安全质量检测学报, columnName=专题：食品中有毒有害物质分析与监测, runingTitle=null, highlight=null, articleAbstract=

目的 建立豆类食品中大豆凝集素的酶联免疫吸附法检测方法。方法 样品经洗净后, 加入磷酸盐缓冲溶液充分匀浆后, 经离心提纯后, 收集上清液经酶联免疫吸附法测定, 用酶标仪定量。结果 在0~400 μg/mL的线性范围内, 所得大豆凝集素的线性方程为Y=0.0021X+0.0528, r=0.9998, 该方法的检出限为125 mg/kg, 回收率在99.17%~101.20%之间, 相对标准偏差在1.27%~2.24%。结论 该方法操作简便, 特异性好, 抗原浓度与OD450值之间呈正比, 适用于基层疾控中心对大豆凝集素中毒事件的检测。

, correspAuthors=张颖茜, authorNote=null, correspAuthorsNote=
* 张颖茜(1987—), 女, 主管技师, 主要研究方向为理化检验。Email:
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陈峰(1978—), 男, 主任技师, 主要研究方向为理化检验工作。E-mail:

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陈峰(1978—), 男, 主任技师, 主要研究方向为理化检验工作。E-mail:

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陈峰(1978—), 男, 主任技师, 主要研究方向为理化检验工作。E-mail:

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Double antibody sandwich enzyme-linked immunosorbent assay for detecting egg white protein in fish mince products[J]. Journal of Food Safety & Quality, 2023, 14(6): 190-197., articleTitle=Double antibody sandwich enzyme-linked immunosorbent assay for detecting egg white protein in fish mince products, refAbstract=null)], funds=[Fund(id=1167158577090273348, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881497364541909, awardId=BE2023772, language=CN, fundingSource=江苏省重点研发计划项目(社会发展)(BE2023772), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1167158574414307343, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881497364541909, xref=null, ext=[AuthorCompanyExt(id=1167158574418501648, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881497364541909, companyId=1167158574414307343, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1. 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Accuracy and precision recovery rate experiment

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编号 质量浓度/(μg/mL) 测得值平均
质量浓度
/(μg/mL)		 平均质量浓度
/(μg/mL)		 RSDs/%
1 50 50.3、50.6、48.9、51.9、48.2、51.2 50.2 1.27
2 100 98.0、100.6、100.9、102.0、100.8、102.1 100.7 1.35
3 400 399.5、398.3、400.6、403.7、402.8、400.9 400.6 2.24
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准确度、精密度实验

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编号 质量浓度/(μg/mL) 测得值平均
质量浓度
/(μg/mL)		 平均质量浓度
/(μg/mL)		 RSDs/%
1 50 50.3、50.6、48.9、51.9、48.2、51.2 50.2 1.27
2 100 98.0、100.6、100.9、102.0、100.8、102.1 100.7 1.35
3 400 399.5、398.3、400.6、403.7、402.8、400.9 400.6 2.24
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Recovery rate experiment with added standards

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编号 底液质量
浓度/(μg/mL)		 加标质量
浓度
/(μg/mL)		 测得值平均
质量浓度
/(μg/mL)		 回收率
/%
1 0.0 50 50.12 100.24
2 0.0 100 101.20 101.20
3 0.0 400 396.70 99.17
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加标回收率实验

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编号 底液质量
浓度/(μg/mL)		 加标质量
浓度
/(μg/mL)		 测得值平均
质量浓度
/(μg/mL)		 回收率
/%
1 0.0 50 50.12 100.24
2 0.0 100 101.20 101.20
3 0.0 400 396.70 99.17
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酶联免疫吸附法检测芸豆中毒相关的大豆凝集素
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陈峰 1 , 钟恩德 2 , 明小天 1 , 张颖茜 1, * , 张婧婧 3
食品安全质量检测学报 | 专题：食品中有毒有害物质分析与监测 2025,16(10): 186-191
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食品安全质量检测学报 | 专题：食品中有毒有害物质分析与监测 2025, 16(10): 186-191
酶联免疫吸附法检测芸豆中毒相关的大豆凝集素
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陈峰1 , 钟恩德2, 明小天1, 张颖茜1, * , 张婧婧3
作者信息
  • 1.南通市疾病预防控制中心, 南通 226007
  • 2.青岛市疾病预防控制中心, 青岛 266071
  • 3.江苏省疾病预防控制中心, 南京 210009

    • 陈峰(1978—), 男, 主任技师, 主要研究方向为理化检验工作。E-mail:

通讯作者:

* 张颖茜(1987—), 女, 主管技师, 主要研究方向为理化检验。Email:
Enzyme-linked immunosorbent assay for detecting soybean agglutinin associated with kidney bean poisoning
Feng CHEN1 , En-De ZHONG2, Xiao-Tian MING1, Ying-Qian ZHANG1, * , Jing-Jing ZHANG3
Affiliations
  • 1. Nantong Center for Disease Control and Prevention, Nantong 226007, China
  • 2. Qingdao Center for Disease Control and Prevention, Qingdao 266071, China
  • 3. Jiangsu Provincial Center for Disease Control and Prevention, Nanjing 210009, China
出版时间: 2025-05-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250226001
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摘要
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目的 建立豆类食品中大豆凝集素的酶联免疫吸附法检测方法。方法 样品经洗净后, 加入磷酸盐缓冲溶液充分匀浆后, 经离心提纯后, 收集上清液经酶联免疫吸附法测定, 用酶标仪定量。结果 在0~400 μg/mL的线性范围内, 所得大豆凝集素的线性方程为Y=0.0021X+0.0528, r=0.9998, 该方法的检出限为125 mg/kg, 回收率在99.17%~101.20%之间, 相对标准偏差在1.27%~2.24%。结论 该方法操作简便, 特异性好, 抗原浓度与OD450值之间呈正比, 适用于基层疾控中心对大豆凝集素中毒事件的检测。

酶联免疫吸附法  /  芸豆  /  中毒  /  大豆凝集素

Objective To establish an enzyme-linked immunosorbent assay method for detecting soybean agglutinin in legume foods. Methods After washed the sample, added phosphate buffer solution and homogenize it thoroughly. After centrifugation and purification, collected the supernatant and determine it by enzyme-linked immunosorbent assay. Quantify it used an enzyme-linked immunosorbent assay reader. Result In the linear range of 0-400 μg/mL, the linear equation of soybean agglutinin obtained was Y=0.0021X+0.0528, r=0.9998. The detection limit of this method was 125 mg/kg, the recovery rate was between 99.17% and 101.20%, and the relative standard deviation was between 1.27%-2.24%. Conclusion This method is easy to operate, has good specificity, and the antigen concentration is directly proportional to the OD450 value. Applicable to the detection of soybean agglutinin poisoning incidents by grassroots disease control centers.

enzyme-linked immunosorbent assay method  /  kidney bean  /  poisoning  /  soybean agglutinin
陈峰, 钟恩德, 明小天, 张颖茜, 张婧婧. 酶联免疫吸附法检测芸豆中毒相关的大豆凝集素. 食品安全质量检测学报, 2025 , 16 (10) : 186 -191 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250226001
Feng CHEN, En-De ZHONG, Xiao-Tian MING, Ying-Qian ZHANG, Jing-Jing ZHANG. Enzyme-linked immunosorbent assay for detecting soybean agglutinin associated with kidney bean poisoning[J]. Journal of Food Safety & Quality, 2025 , 16 (10) : 186 -191 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250226001
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0 引言
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大豆凝集素(soybean agglutinin, SBA)是一种植物凝集素和抗营养因子,食入未煮熟的豆类食品, 会致使红细胞彼此黏聚, 增加血液黏稠度, 从而诱发血栓[1-2]。根据美国食品和药物管理局的相关通知, 4颗生芸豆就有可能导致严重的恶心、呕吐和腹泻症状。大豆凝集素的常用检测方法有植物性代谢产物大豆凝集素测定酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)和转基因生物及其产品食用安全检测抗营养因子大豆中凝集素检测方法液相色谱-串联质谱法、聚合酶链式反应、多种食用豆科植物毒性试验方法联合皂甙定性检测和红细胞凝集实验等[3-4]。液相色谱-串联质谱法能够对大豆凝集素的测量进行准确定量, 利用标准肽一和标准肽二的保留时间和二级谱图(或者子离子)进行定性[5-7], 但其有检测仪器、试剂昂贵, 分析过程耗时长等缺点。红细胞凝集实验操作条件要求高、且程序较复杂, 无法满足突发快速检测和大批日常检测的要求, 而ELISA法作为分析化学领域的检测方法之一, 具有选择性强, 检测时间短, 成本低, 操作方便等优势。
鉴于此, 本研究采用双抗体夹心ELISA法(见图1)建立了一种ELISA法检测芸豆中毒中的大豆凝集素方法, 适用于基层疾控中心对大豆凝集素中毒事件的检测, 并在450 nm波长下对光密度(optical density, OD)值进行测定。以期为现场处理大豆凝集素的食物中毒事件提高科学依据。
1 材料与方法
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1.1 仪器、试剂与材料
LA204电子分析天平(精度0.1 mg, 常熟百灵电子天平公司); IMARK酶标仪(伯乐-550型、450 nm波长滤光片, 美国BIO-RAD公司); MultIFUGE X1R PrimoB高速冷冻离心机(美国Thermo Fisher Scientific公司); HH-2电热恒温水浴锅(国华仪器制造有限公司); AdvantageA10 Milli-QA超纯水器(美国Millipiore公司)。
大豆凝集素ELISA试剂盒(批号: A112664-48T)、大豆凝集素标准品(批号: A112664-48T)(上海抚生实业有限公司); 磷酸盐缓冲溶液(北京兰杰柯科技有限公司); 实验用水都为超纯水。
样品来源某食品店集中供餐采购的芸豆。
1.2 实验方法
1.2.1 原理
采用双抗体夹心ELISA法原理: 用待检测的大豆凝集素与已经包被在聚苯乙烯板上的大豆凝集素抗体充分反应, 再加入HRP标记的大豆凝集素抗体, 形成抗体-抗原-酶标记抗体复合物, 通过洗板, 洗去多余游离抗体、抗原及杂质, 再加入底物四甲基联苯胺显色, 使测定方法达到较高的敏感度, 反应颜色的深浅与待测的大豆凝集素含量成正比。记录酶标仪在450 nm波长处OD, 配制系列标准溶液后制作相应的标准曲线, 大豆凝集素浓度与OD450值之间呈正比, 通过标准曲线可获得样品中大豆凝集素的浓度。
1.2.2 仪器工作条件
仪器工作条件为酶标仪450 nm波长, HH-2电热恒温水浴锅进行水浴操作。
1.2.3 样品的收集、保存
使用干燥、清洁的容器采集疑似中毒的样品, 短期保存的样本应放置在4 ℃低温冰箱中, 保存时间一般不超过一周, 实验时要避免反复冻融。长期保存的样本应放置在-20 ℃或更低温度的冰箱中, 以避免样品变质或产生污染。样本应放置在清洁干燥的区域, 并保持密封状态, 避免外界污染, 确保样品的质量和稳定性。
1.2.4 样品预处理
将标本室温放置30 min, 准确称取留存的芸豆食物样品1 g置于15 mL离心管中, 加入5.0 mL 磷酸盐缓冲溶液(pH 7.4), 手工或匀浆器将标本充分混匀, 在4 ℃低温高速离心机以2000~3000 r/min离心20 min, 收集上清液[8], 分装若干份小瓶待测定-20 ℃下存放, 并尽快完成测定[9-11]。如果是富含大豆凝集素的样品, 需稀释合适倍数后再检测[12-15]。
1.2.5 标准曲线的绘制
(1)标准品的稀释
先吸取150 μL试剂盒中800 mg/L的大豆凝集素溶液加入150 μL标准品稀释液中, 依梯次逐级稀释配制成0、25、50、100、200、400 μg/mL的大豆凝集素标准溶液系列; 加到酶标板中。
(2)加样
准确吸取50 μL大豆凝集素标准溶液加进6个包被大豆凝集素抗体微孔板里, 待测样品孔同样加入50 μL(样品10 μL与样品稀释液40 μL混匀), 标准与样品加样时移液枪尽量悬浮在微孔上方, 小心吹打混匀后, 轻磕96孔板, 确保微孔壁上无液体。
(3)孵育
封板膜密封后, 37 ℃孵育30 min。
(4)配液
把浓缩洗涤液稀释30倍待用。
(5)洗涤
撕掉封板膜后, 弃去微孔中的游离的大豆凝集素或大豆凝集素抗体, 倒扣96孔板拍打, 所用的微孔内加入上述洗涤液, 反应0.5 min左右后弃去液体, 如此洗涤操作重复4次后, 拍干。
(6)加酶
除空白孔, 其他孔均加入HRP标计大豆凝集素抗体。
(7)孵育、洗涤
操作过程同1.2.5(3)和1.2.5(5)。
(8)显色
在标准系列孔与样品微板孔中分别缓缓注入50 μL显色剂A与加入50 μL显色剂B, 混匀, 孵育箱中37 ℃左右催化显色反应10 min以上, 标准系列溶液呈蓝色。
(9)终止
在所有微孔中均加50 μL稀硫酸终止液, 显色反应终止, 标准系列颜色转为黄色。
(10)测定
用空白标准品溶液的吸光度调零, 检测波长为450 nm, 分别记录所有微板孔中的吸光度(OD值)[16-17]。 注意测定标准系列与样品中大豆凝集素含量应在加终止液后15 min内进行。以不同浓度大豆凝集素标准品为横坐标(X, μg/mL)、相对光密度OD所测数值作为纵坐标(Y), 构建标准曲线, 进而计算出该曲线的回归方程式以及对应的相关系数, 在0~400 μg/mL的线性范围内, 所得大豆凝集素的线性方程为Y=0.0021X+0.0528, r=0.9998。流程图见图2。
1.2.6 样品的测定
样品测定与标准曲线测定同时进行, 在标记好的待测样品孔中, 操作时首先注入40 μL的样品稀释液, 随后再注入10 μL的样品提取液, 加样时移液枪头尽量不触碰孔壁, 加好样品液之后轻轻混匀晃动, 注意不要晃出管外, 按照标准曲线绘制步骤测定样品空白、样品的OD值, 以样品减去试剂空白的OD值后, 由绘制的标准曲线计算样品中大豆凝集素的浓度, 见公式(1)。
$X=\frac{\left(\rho_{\text {样 }}-\rho_{\text {空 }}\right) \times V \times a \times 1000}{m \times 1000}$
式中: X为大豆凝集素的浓度, mg/kg; ρ样为样品提取液中大豆凝集素的质量浓度, μg/L; ρ空为空白液中大豆凝集素的质量浓度, μg/L; V为样品处理液的体积, mL; a为样品的稀释倍数; m为样品重量, g。
1.3 数据处理
记录标准品和样本的吸光度(OD值), 减去空白孔(无样本或试剂的对照)的OD值, 消除非特异性信号, 将OD值导入Microsoft Excel 2021, 剔除异常值, 同一样本在同一板上重复检测≥3次, 评估操作误差, 通过上述方法, 系统完成从数据采集到结果可视化的全流程分析, 确保实验结果的可靠性与科学性。
2 结果与分析
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2.1 测定条件优化
2.1.1 磷酸盐缓冲液pH选择
通过实验发现, pH 7.4的磷酸盐缓冲液的pH与大豆凝集素在人体内部分环境的pH相似, 较为稳定, 不会对细胞和分子的生理功能产生影响; 且缓冲能力较强, pH 7.4的磷酸盐缓冲液在这个pH下具有较强的缓冲能力, 可以有效地维持溶液的pH稳定。反应体系中大豆凝集素浓度和缓冲溶液pH对实验影响程度较大, 是完善ELISA检测标准化程序的关键因素[18-21]。
2.1.2 抗原、抗体的最适工作浓度的确定
将大豆凝集素的标准溶液0~400 μg/mL范围内与酶标孔中的捕获抗体捕捉样品中的游离抗原, 与检测抗体形成夹心复合物, 结合酶标二抗, 通过显色反应判定样品中抗原物质的浓度。酶标仪通常要求在18~25 ℃的室温下使用[22-26], 测得标准曲线相关系数r=0.9998。
2.1.3 孵育、洗涤时间的选择
大豆凝集素标准溶液与样品溶液各50 μL保持环境温度在20 ℃左右, 孵育时间为30 min, 孵育温度为37 ℃左右和显色时间为10 min、洗涤至少4次, 用力甩干, 不能有残余液体, 用酶标仪显色测定效果较好[27-29]。
2.1.4 洗涤液选择
使用试剂盒配套的浓缩洗涤液30倍稀释后使用, 以确保实验结果的准确性和一致性‌, 如果试剂盒内的洗涤液用完, 建议使用同一批号的洗涤液; 不建议混用不同系列项目的洗涤液, 不同项目的洗涤液成分可能不同[30-31], 可能会影响实验结果。
2.2 线性范围、回归方程与检出限
大豆凝集素在0~400 μg/mL的线性范围内, 线性良好, 其回归方程为Y=0.0021X+0.0528, 相关系数r=0.9998, 此次生物试剂盒的检出限为125 mg/kg。
2.3 准确度、精密度实验
分别向超纯水(空白样)不同的样品孔中加入大豆凝集素标准溶液, 配制成50、100、400 μg/mL 3个浓度点, 每个浓度均测定6次, 进行批间精密度计算, 计算测定结果的相对标准偏差(relative standard deviation, RSD), RSD在1.27%~2.24%。见表1。
2.4 加标回收率实验
为验证ELISA实验方法检测大豆凝集素的准确性, 需要对大豆凝集素做加标回收实验。 根据大豆凝集素标准曲线的OD值确定3个加标浓度。分别向超纯水(空白样)不同的样品孔中加入大豆凝集素标准溶液, 配制成50、100、400 μg/mL 3个质量浓度点, 每个浓度均测定6次, 用测定平均值减去空白值后计算大豆凝集素的加标回收率。做一组空白, 加标回收率在99.17%~101.20%之间, 见表2。
2.5 样品测定
由某食品店集中供餐引起的一起芸豆中毒事件, 经调查芸豆初加工洗净后入沸水焯水约30 s捞出, 入油锅加蒜泥调味翻炒1 min左右出锅。调查员现场查看留样食品, 蒜泥芸豆颜色青绿, 指掐感觉较为生硬。部分患者自述食用蒜泥芸豆感觉比较生未煮熟。从采集的留样食品蒜泥芸豆检出大豆凝集素, 对四季豆样品进行多次测定, 测得均值为225 mg/kg。食物安全管理机构, 包括联合国粮食及农业组织/世界卫生组织食物添加剂联合专家委员会等, 并未就植物血球凝集素进行评估, 未订定用作风险评估的健康参考值。
3 讨论与结论
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使用电热恒温水槽时要加入适量的水, 通常是加至离槽口约5 cm的位置, 避免水位过低导致加热元件露出水面造成损坏; 校准温度计以确保实验时的温度读数准确, ‌‌使用过程中的注意事项, 根据实验需求设置37 ℃, 避免过高或过低的温度造成实验误差或损坏实验物品; 缓慢升温, 不要将温度设定过高后突然打开加热器, 以防水温急剧上升导致水溢出或设备过热; 在加热过程中注意及时补水, 避免因水位过低而导致设备过热‌, 操作时需小心。
双抗体夹心ELISA法在对豆类食品中大豆凝集素的实验中, 准确性和灵敏度都较稳定。ELISA测定大豆凝集素的报道不多, 本方法利用双抗体夹心ELISA法在准确度、精密度、回收率实验中RSD在1.27%~2.24%, 加标回收率在99.17%~101.20%之间。与前期研究结果一致, 庞杏豪[9]作出大豆凝集素免疫学快速检测方法的研究。就成分相对单一的豆类食品而言, 夹心ELISA法在检测其中大豆凝集素时更有优势, 能更为精准高效地完成任务; 而那些经过深度加工、在较为严苛加工条件下制成的食品, 由于成分复杂、结构改变等因素影响, 竞争ELISA法就成了测定其内部大豆凝集素含量的优选策略。在此基础上, 本研究初步搭建起了双抗体夹心ELISA法检测大豆凝集素的方法, 通过实际中毒样品的检测, 结果表明, 双抗体夹心ELISA法适用于中毒样品准确的检测。在进行加标回收实验中, 有复杂基质效应的存在, 中毒食物样品越是复杂, 回收率就越低, 可能是因为食物复杂基质组分之间的相互作用来影响大豆凝集素的检测。‌‌‌
本研究中ELISA法检测大豆凝集素的方法, 分析时间短, 重现性较好, 操作简便, 检测费用成本低等优势。由于其携带方便、检测速度快, 很适合基层疾控中心对疑似大豆凝集素中毒事件进行定性、定量分析, ELISA法检测大豆凝集素必备条件允许可应用于现场检测中毒样品。缺点是有效期只有半年。此次在检测大豆凝集素的同时, 实验人员要掌握酶标仪使用的基础操作, 保持孵育箱的温度平衡, 不同样品处理的提取体积。在预防大豆凝集素中毒的同时, 餐饮服务单位要严格执行食品安全管理制度, 保持操作环境卫生整洁, 食物制作应烧熟煮透, 四季豆应炒或焖至颜色全变, 无豆腥味再食用。加强对食品经营者食品卫生及食品安全知识培训, 严格执行食品安全法有关规定, 及早采取预防措施, 严格执行留样制度。通过公众号向社会进行芸豆中毒的知识宣传, 增强群众自我保护意识, 确保自身的食用安全。本研究运用双抗体夹心ELISA法测定大豆凝集素实验适合基层疾病预防控制中心定性定量使用。
基金
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  • 江苏省重点研发计划项目(社会发展)(BE2023772)
文献
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2025年第16卷第10期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250226001
  • 接收时间：2025-02-26
  • 首发时间：2025-07-15
  • 出版时间：2025-05-25
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  • 收稿日期：2025-02-26
基金
江苏省重点研发计划项目(社会发展)(BE2023772)
作者信息
    1.南通市疾病预防控制中心, 南通 226007
    2.青岛市疾病预防控制中心, 青岛 266071
    3.江苏省疾病预防控制中心, 南京 210009

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* 张颖茜(1987—), 女, 主管技师, 主要研究方向为理化检验。Email:
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
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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