粮谷中呕吐毒素管式化学发光免疫分析法的研究与应用

王毅谦; 陈君义; 龙云凤; 陆慧媛; 方云涛; 于洋

doi:10.13386/j.issn1002-0306.2017.03.048

粮谷中呕吐毒素管式化学发光免疫分析法的研究与应用

Investigation and application of a tubular Chemiluminiscence immunoassay on detection of Deoxynivalenol

摘要

摘要: 目的:建立一种准确度高、重复性优的快速检测粮谷中呕吐毒素（DON）的管式化学发光免疫分析法（CLIA）。方法:本研究基于抗原抗体的特异性反应,选择磁性微粒为固相载体,通过优化磁微粒的稀释度、反应时间等工作条件,建立了高准确度、重复性良好的DON-CLIA。结果:本研究建立的DON-CLIA的检测范围为5¹85 ng/m L;最低检测限为3.82 ng/m L（样本检测限为0.076μg/g）;准确度相对偏差为6.64%,重复性变异系数为2.32%,均小于10%;DON的添加回收率在90%¹10%之间。结论:本研究建立的DON-CLIA的检测范围、检测限、准确度、重复性、回收率,均符合实际工作的要求,是一种快速、高效、准确的化学发光免疫分析法,适用于大量样本的DON含量的测定。

Abstract: Objective: To establish a high accurate and excellently repeatable tubular Chemiluminiscence immunoassay( CLIA)for rapid detection of Deoxynivalenol( DON) in cereals.Method: Based on the specific antigen- antibody reaction,magnetic particles were used as the solid phase carrier. The operating conditions,such as the concentration of magnetic particles and reaction time,were optimized.A high accurate tubular CLIA for quantifying of DON( DON- CLIA) in cereals was established.Result: This DON- CLIA showed an excellent performance in linear with range of 5~185 ng / m L.The limit of detection( LOD)was 3.82 ng / m L.Both relative deviation of accuracy( 6.64%) and coefficient of variation of repeatability( 2.32%) were less than 10%.The percent recovery of DON was 90% ~110%.Conclusion: All performance parameters of DON- CLIA,such as detection range,LOD,accuracy,repeatability and recovery,areaccorded with requirement of DON detection. This DON- CLIA,with high sensitivity and specificity,is suitable for quantitative determination of DON in cereals.