基于间接竞争原理的流式微球技术快速检测麦芽中赭曲霉毒素A

建立了一种快速、灵敏检测中药麦芽中赭曲霉毒素A（Ochratoxin A,OTA）含量的间接竞争流式微球方法。麦芽样品经60%甲醇/PBS提取,加入20%甲醇/PBS溶液稀释5倍后,离心取上清液制备样品。在编码荧光微球上偶联牛血清白蛋白-OTA（BSA-OTA）复合物,与样品中OTA竞争结合抗OTA特异性抗体,然后加入FITC-IgG孵育,离心洗涤后,用流式细胞仪检测微球表面的平均荧光强度,实现样品中OTA的准确定性定量测定。本方法检测OTA的半数抑制浓度IC_（50）为1.20 ng/mL,相关系数R^2=0.9892,检出限（IC_（10））为0.12 ng/mL,加样回收率为93.9%~...

Full description

Saved in:
Bibliographic Details
Published in分析化学 Vol. 44; no. 4; pp. 625 - 632
Main Author 肖昌彬 刘秋桃 豆小文 杨美华 孔维军 万丽
Format Journal Article
LanguageChinese
Published 中国医学科学院北京协和医科大学,药用植物研究所,北京100193%中国医学科学院北京协和医科大学,药用植物研究所,北京100193 2016
辽宁医学院药学院,锦州121001%中国医学科学院北京协和医科大学,药用植物研究所,北京100193%成都中医药大学药学院,成都,611137
成都中医药大学药学院,成都611137
Subjects
快速检测
流式微球技术
赭曲霉毒素A
间接竞争
麦芽
流式微球技术
间接竞争
麦芽
赭曲霉毒素A
Indirect competition
Cytometric bead array
Malt
快速检测
Ochratoxin A
Rapid detection
Online AccessGet full text
ISSN0253-3820
DOI10.11895/j.issn.0253-3820.151007

Cover

More Information
Summary:建立了一种快速、灵敏检测中药麦芽中赭曲霉毒素A（Ochratoxin A,OTA）含量的间接竞争流式微球方法。麦芽样品经60%甲醇/PBS提取,加入20%甲醇/PBS溶液稀释5倍后,离心取上清液制备样品。在编码荧光微球上偶联牛血清白蛋白-OTA（BSA-OTA）复合物,与样品中OTA竞争结合抗OTA特异性抗体,然后加入FITC-IgG孵育,离心洗涤后,用流式细胞仪检测微球表面的平均荧光强度,实现样品中OTA的准确定性定量测定。本方法检测OTA的半数抑制浓度IC_（50）为1.20 ng/mL,相关系数R^2=0.9892,检出限（IC_（10））为0.12 ng/mL,加样回收率为93.9%~97.4%,RSD〈3.6%。16份实际麦芽样品中检测到2个阳性样品,OTA的最高含量为3.83μg/kg,未超出欧盟的OTA限量标准,与液相色谱-串联质谱（LC-MS/MS）法检测结果相一致。本方法简单、快速、灵敏、可靠,可拓展用于其它复杂基质中多种真菌毒素的定性与定量检测。
Bibliography:Cytometric bead array; Indirect competition; Malt; Ochratoxin A; Rapid detection
22-1125/O6
A cytometric bead array（ CBA） method based on indirect competition mode was developed for the sensitive and rapid detection of ochratoxin A（ OTA） in malt. The malt samples were extracted by 60%methanol / PBS and the extracts were diluted five times by 20% methanol / PBS,then the supernatant was collected to prepare sample solution for analysis. The fluorescence microsphere surface was labeled with bovine serum albumin-OTA（ BSA-OTA） to compete with OTA in the sample for anti-OTA monoclonal antibody（ mA b）. Then FITC-Ig G was added to bind with the captured mA b on the microsphere. Finally,the mean fluorescence intensity of FITC on the surface of microsphere was detected by a BD FACSCalibur analyzer for accurate qualitative and quantitative analysis of OTA. The results showed that the half inhibitory concentration（ IC_（50）） was 1. 20 ng /mL with a correlation coefficient（ R~2） of 0. 9892,and the limit of detection（ LOD） fo
ISSN:0253-3820
DOI:10.11895/j.issn.0253-3820.151007