文章摘要
潘明飞;李诗洁;郭丹丹;王俊平;王硕.间接竞争酶联免疫分析方法检测花生中黄曲霉毒素B1[J].中国食品学报,2019,19(9):255-261
间接竞争酶联免疫分析方法检测花生中黄曲霉毒素B1
Determination of Aflatoxin B1 in Peanut by Indirect Competitive Enzyme-linked Immunosorbent Assay
  
DOI:
中文关键词: 黄曲霉毒素B1  间接竞争酶联免疫分析方法  花生
英文关键词: aflatoxin B1  ic-ELISA  peanut
基金项目:国家自然科学基金项目(31972147);天津市自然科学基金项目(17JCQNJC14800);天津市高等学校基础科研项目(2017KDZD01)
作者单位
潘明飞;李诗洁;郭丹丹;王俊平;王硕 教育部食品安全与营养重点实验室天津科技大学天津300457 
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中文摘要:
      目的:针对花生中强毒性污染物黄曲霉毒素B1(AFB1)建立了一种准确、灵敏的间接竞争酶联免疫分析方法(ic-ELISA)。方法:制备了AFB1包被抗原,包被量为0.1 μg/孔;抗体稀释64 000倍;选用0.1%脱脂乳粉为方法封闭液,0.01 mol/L磷酸盐缓冲液(PBS,pH 7.4)为样品稀释液;结果:在最优的试验条件下,建立的ic-ELISA对目标物AFB1的检测线性范围为0.0097~0.088 μg/L(R2 = 0.999);灵敏度(IC50)和检出限(IC15)分别达到0.027 μg/L和0.0066 μg/L。板内变异系数(n=6)为0.29%~16.9%,板间变异系数(n=6)为0.22%~21.19%。在花生样品中AFB1的检测灵敏度(IC50)为1.04 μg/kg,回收率为96.67%~106.51%。结论:本研究建立的ic-ELISA方法操作相对简便,检测灵敏度高、准确性好,可稳定地用于花生样品中AFB1污染物的定量分析检测。
英文摘要:
      Purpose: An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for detection of aflatoxin B1(AFB1) in peanut was developed. Methods: The AFB1 antigen was prepared, and coated in 96 well plates with the content of 0.1 μg/well; antibody was diluted 64 000 times; the 0.1% skimmed milk powder solution was selected as the blocking buffer, 0.01 mol/L phosphate buffer solution (PBS, pH 7.4) was used as dilution buffer to establish the proposed ic- ELISA detection method. Results: The limit of detection limit (LOD, IC15) of this ic-ELISA assay was 0.0066 μg/L; the sensitivity (IC50) was 0.027 μg/L; and the linear range was 0.0097-0.088 μg/L (R2 = 0.999). The intra-assay variation coefficient (n=6) range was 0.29%-16.9%, and the inter-assay variation coefficient (n=6) range was 0.22%-21.19%. The sensitivity(IC50) in peanut sample was 1.04 μg/kg, and the recovery range was 96.67%-106.51%. Conclusion: The ic-ELISA assay established in this study was simple, sensitive and accurate and can be used for quantitative determination of AFB1 in peanut samples.
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