基于核酸适配体的侧流层析技术同步检测赭曲霉毒素A和黄曲霉毒素B1

Synchronous detection of ochratoxin A and aflatoxin B1 by lateral flow chromatography based on nucleic acid aptamers

中文关键词:  赭曲霉毒素A  黄曲霉毒素B1  适配体  试纸条  花生和葡萄干

英文关键词:ochratoxin A  aflatoxin B1  aptamer  test strip  peanut and raisin

中文摘要:

      目的 建立基于适配体互补链同时检测赭曲霉毒素A(ochratoxin A, OTA)和黄曲霉毒素B1(aflatoxin B1, AFB1)2种真菌毒素的荧光试纸条。方法 通过优化2种适配体的浓度以及缓冲体系的pH值, 提高同步检测OTA和AFB1的灵敏度和准确性。结果 OTA和AFB1的T线(TO和TA)和C线荧光强度比值与对应真菌毒素的浓度对数具有良好的线性关系, 线性范围为0.5~50 ng/mL, 相关系数r2分别为0.9887和0.9910, 检出限低至0.51和0.38 ng/mL。通过对花生和葡萄干进行加标回收和实际样品检测, 试纸条检测的OTA和AFB1的回收率分别为82.06%~109.69%和83.34%~110.06%, 相对标准偏差(relative standard deviation, RSD)为1.89%~8.17%, 检测结果与高效液相色谱-串联质谱法(high performance liquid chromatography-mass spectrum/mass spectrometry, HPLC-MS/MS)一致。结论 该生物传感器可在20 min内实现花生和葡萄干等样品中OTA和AFB1的同步快速检测, 并具有成本低、检测速度快和易于操作等优点, 可为多种真菌毒素的同步快检提供技术支撑。

英文摘要:

      Objective To establish a fluorescent test strip based on the aptamer complementary strand for the simultaneous detection of 2 mycotoxins, ochratoxin A (OTA) and aflatoxin B1 (AFB1). Methods The sensitivity and accuracy of simultaneous detection of OTA and AFB1 were improved by optimizing the concentration of the 2 aptamers and the pH value of the buffer system. Results The T line (TO and TA) and C line fluorescence intensity ratios of OTA and AFB1 showed a good linear relationship with the logarithm of the concentration of the corresponding mycotoxin in the linear range of 0.5?50 ng/mL, with the correlation coefficients r2 of 0.9887 and 0.9910, respectively. The detection limits were as low as 0.51 and 0.38 ng/mL. The recoveries of OTA and AFB1 detected by test strips were 82.06%?109.69% and 83.34%?110.06%, respectively, and the relative standard deviations were 1.89%?8.17%. The detection results were consistent with those of high performance liquid chromatography-tandem mass spectrometry. Conclusion The biosensor can realize synchronous and rapid detection of OTA and AFB1 in peanut, raisin and other samples within 20 min, has the advantages of low cost, high detection speed, easy operation and the like, and can provide technical support for synchronous and rapid detection of a plurality of mycotoxins.