Development and comparison of three diagnostic immunoassay formats for the detection of azoxystrobin
Development and comparison of three diagnostic immunoassay formats for the detection of azoxystrobin
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DOI：
10.1021/jf052424o
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发表时间：
2006-02-08
影响因子：
6.1
通讯作者：
Hall, JC
中科院分区：
文献类型：
--
作者：
Furzer, GS;Veldhuis, L;Hall, JC
目前公认的检测嗜球果伞素类杀菌剂嘧菌酯的方法涉及劳动密集型有机溶剂萃取和气相色谱分析。三种诊断测定形式，即，酶联免疫吸附试验（ELISA），荧光偏振法（FP），时间分辨荧光法（TR-FIA），并进行了比较方面的检测和定量嘧菌酯在葡萄提取物和河流，湖泊和井水样品。这三种测定形式不需要初始样品提取，并且不受任何检测的环境基质的影响，并且每种测定形式的线性工作范围为0-400 pg/mL。用于每个免疫测定的多克隆抗体是特定的嘧菌酯，也就是说，观察到的其他农药的最高交叉反应性为5.7%。免疫测定的检测限相似，分别为3（ELISA）、46（FP）和28（TR-FIA）pg/mL，IC 50值也相似，分别为306、252和244 pg/mL。三种免疫检测方法的劳动强度较低，灵敏度约为气相色谱法的100倍。虽然三种形式在性能方面相当，但荧光偏振测定是劳动强度最小的，并且需要最少的时间来进行。
The currently accepted method of detection for azoxystrobin, a strobilurin fungicide, involves a labor-intensive organic solvent extraction and gas chromatography analysis. Three diagnostic assay formats, i.e., enzyme-linked immunosorbent assay (ELISA), fluorescence polarization (FP), and time-resolved fluorescence (TR-FIA), were developed and compared with regard to detection and quantification of azoxystrobin in grape extract and river, lake, and well water samples. These three assay formats require no initial sample extraction and were not affected by any of the environmental matrices tested, and each had a linear working range of 0-400 pg/mL. The polyclonal antibodies used for each of the immunoassays were specific to azoxystrobin; that is, the highest cross-reactivity to other pesticides observed was 5.7%. The limits of detection of the immunoassays were similar at 3 (ELISA), 46 (FP), and 28 (TR-FIA) pg/mL, as were the respective IC50 values of 306, 252, and 244 pg/mL. Each of the three immunoassays developed was less labor-intensive and approximately 100-fold more sensitive than the gas chromatographic method. While the three formats were comparable in terms of performance, the fluorescence polarization assay was the least labor-intensive and required the least time to perform.