基于金纳米花免疫层析法同时检测大米中呕吐毒素和黄曲霉毒素B1含量

Simultaneous detection of deoxynivalenol and aflatoxin B1 content in rice based on gold nanoflowers immunochromatography assay

中文关键词:  金纳米花  多重免疫层析法  黄曲霉毒素B1  呕吐毒素

英文关键词:gold nanoflowers  multiplex immunochromatography  aflatoxin B1  deoxynivalenol

中文摘要:

      目的 谷物中多种真菌毒素共污染问题已成为食品安全的重大挑战,为保障粮食安全,本研究建立了同时快速检测大米中呕吐毒素(deoxynivalenol,DON)和黄曲霉毒素B1(aflatoxin B1,AFB1)的方法。 方法 通过制备金纳米花(gold nanoflowers, AuNFs),并优化标记pH值、抗体用量、探针用量和探针混合比例等关键因素,评价AuNFs免疫层析试纸条对DON和AFB1同步检测性能。 结果 结果表明,该方法在标记pH为7.0、抗体用量为12 μg和9 μg、探针用量2.5 μL及DON-mAb和AFB1-mAb探针混合比1:2检测条件下,对DON和AFB1标准品的可视化检出限分别为0.5 ng/mL、0.2 ng/mL,且对四种常见真菌毒素具有良好的特异性。将该方法应用于大米样品检测时,DON和AFB1检测限分别为5 ng/g和1 ng/g。 结论 因此,研究构建的AuNFs双重免疫层析试纸条具有操作便捷、快速、灵敏等特点,符合谷物真菌毒素检测的国家限量标准,为实现现场快速大批量筛查大米中DON、AFB1提供了新的思路。

英文摘要:

      Objective Co-contamination of multiple mycotoxins in cereals has become a major challenge to food safety, and a method for the simultaneous detection of DON and AFB1 in rice has been established to protect food security. Methods The performance of AuNFs immunochromatographic strips for the simultaneous detection of DON and AFB1 was evaluated by preparing gold nanoflowers (AuNFs) and optimizing the several key factors, such as labeling pH, antibody dosage, probe dosage and probe mixing ratio. Results The results showed that the optimizing conditions were 7.0 of labeling pH, 12 μg and 9 μg of antibody dosage, 2.5 μL of probe dosage and 1:2 of mixed ratio of DON-mAb and AFB1-mAb probe, respectively. The visual detection limits of the method for DON and AFB1 standards were 0.5 ng/mL and 0.2 ng/mL, respectively. The built assay exhibited highly specific for four other contaminated mycotoxins. Moreover, the detection limits of DON and AFB1 were 5 ng/g and 1 ng/g in the detection of rice samples, respectively. Conclusion Therefore, the constructed AuNFs dual immunochromatographic test strips in this work realized rapid, sensitive and simple detection of DON and AFB1, meeting the national limit standards for the detection of mycotoxins in cereals, which provide a new idea for the rapid and large-scale screening of two mycotoxins in rice.