同时检测玉米中黄曲霉毒素B1和赭曲霉毒素A的时间分辨荧光免疫层析试纸条的研制

doi:10.7506/spkx1002-6630-20210102-006

食品科学 ›› 2022, Vol. 43 ›› Issue (2): 346-354.doi: 10.7506/spkx1002-6630-20210102-006

同时检测玉米中黄曲霉毒素B1和赭曲霉毒素A的时间分辨荧光免疫层析试纸条的研制

卢迪莎，王序，杨金易，罗林，肖治理

（广东省食品质量安全重点实验室，华南农业大学食品学院，广东广州 510642）

出版日期:2022-01-25 发布日期:2022-01-29
基金资助:
“十三五”国家重点研发计划重点专项（2018YFC1602903）；广州市科技计划项目（201804010403）

Development of a Time-resolved Fluorescence Immunochromatographic Strip for the Simultaneous Detection of Aflatoxin B1 and Ochratoxin A in Corn Samples

LU Disha, WANG Xu, YANG Jinyi, LUO Lin, XIAO Zhili

(Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China)

Online:2022-01-25 Published:2022-01-29

摘要/Abstract

摘要： 研究制备了一种可同时快速定量检测黄曲霉毒素B1（aflatoxin B1，AFB1）和赭曲霉毒素A（ochratoxin A，OTA）的二联时间分辨荧光免疫层析试纸条。采用铕系时间分辨荧光微球分别标记AFB1和OTA单克隆抗体，优化荧光微球活化pH值、标记的抗体浓度、荧光探针使用量、检测线包被原浓度、质控线羊抗鼠IgG浓度、样品前处理条件等因素。结果表明：检测AFB1和OTA的IC50分别为1.58、3.91 μg/kg，线性范围分别为0.26～9.73、1.14～13.29 μg/kg，肉眼检测限分别为3.70、5.55 μg/kg；与黄曲霉毒素类似物交叉率为12%～62%，与赭曲霉毒素B交叉率为58%，与其他真菌毒素无明显交叉反应；选择玉米样品进行添加回收实验，2 种毒素回收率在92%～103%之间，变异系数小于15%；对实际样品的检测与高效液相色谱法和市售胶体金试纸检测结果一致；且该试纸条可在4 ℃稳定保存5 个月以上，稳定性较好。本研究所制备的时间分辨荧光免疫层析试纸条具有灵敏、准确、快速和简便等特点，非常适用于玉米等食品和农产品中AFB1和OTA的现场快速筛查。

关键词: 黄曲霉毒素B1；赭曲霉毒素A；时间分辨；荧光免疫层析试纸条

Abstract: In this study, a time-resolved fluorescence immunochromatographic assay (TRFICA) for detecting aflatoxin B1 (AFB1) and ochratoxin A (OTA) simultaneously was developed. Europium-based time-resolved fluorescent microspheres were used to label anti-AFB1 monoclonal antibody and anti-OTA monoclonal antibody. Various factors such as pH for the activation of fluorescent microspheres, concentration of labeled antibodies, dosage of fluorescent probes, concentration of coating antigens for test line, concentration of goat anti-mouse IgG for quality control line, and sample pretreatment conditions were optimized. Under the optimal conditions, the assay showed half inhibition concentration (IC50) values of 1.58 and 3.91 μg/kg , linear ranges of 0.26–9.73 and 1.14–13.29 μg/kg, and visible limits of detection (vLOD) of 3.70 and 5.55 μg/kg for AFB1 and OTA, respectively. The cross-reactivity rates of this method with aflatoxin analogues and ochratoxin B were 12%–62% and 58%, respectively, without cross-reactivity with other mycotoxins. The recoveries of the two toxins spiked in corn samples were 92%–103%, with coefficients of variation (CV) less than 15%. The results of this method for real samples coincided with those of high performance liquid chromatography (HPLC) and commercial colloidal gold strip. The TRFICA strip was stored at 4 ℃ for more than 5 months with good stability. To sum up, this immunochromatographic strip is sensitive, accurate, rapid and easy to operate, and is very suitable for rapid screening of AFB1 and OTA in food and agricultural products.

Key words: aflatoxin B1; ochratoxin A; time-resolved; fluorescence immunochromatographic strip

中图分类号:

TS201.6

卢迪莎，王序，杨金易，罗林，肖治理. 同时检测玉米中黄曲霉毒素B1和赭曲霉毒素A的时间分辨荧光免疫层析试纸条的研制[J]. 食品科学, 2022, 43(2): 346-354.

LU Disha, WANG Xu, YANG Jinyi, LUO Lin, XIAO Zhili. Development of a Time-resolved Fluorescence Immunochromatographic Strip for the Simultaneous Detection of Aflatoxin B1 and Ochratoxin A in Corn Samples[J]. FOOD SCIENCE, 2022, 43(2): 346-354.

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同时检测玉米中黄曲霉毒素B1和赭曲霉毒素A的时间分辨荧光免疫层析试纸条的研制

Development of a Time-resolved Fluorescence Immunochromatographic Strip for the Simultaneous Detection of Aflatoxin B1 and Ochratoxin A in Corn Samples

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