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首页> 外文期刊>Food analytical methods >Development of an Indirect Competitive ELISA Kit for the Rapid Detection of Benzopyrene Residues
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Development of an Indirect Competitive ELISA Kit for the Rapid Detection of Benzopyrene Residues

机译：快速检测苯并yr残留的间接竞争ELISA试剂盒的开发

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摘要

This paper presents the generation of monoclonal antibodies (mAbs) with high specificity against benzopyrene (BaP) through cell fusion procedures, and the development of mAb-based indirect competitive enzyme-linked immunoabsorbent kit (icELISA kit) to detect BaP using one of these hybridomas (clone 2E12). The calibration curve of BaP-Kit with standard BaP inhibitor was typical sigmoid curve fitted to the four parameters logistic equation with the linear regression equation y = -28.662x + 132.87, the half-maximal inhibition concentration (IC50) of 0.779 mu g/L and the sensitivity of 0.054 mu g/L. The recovery of BaP spiked in beef was between 81.3 and 93.6 %. The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 6 %. Except for minor cross-reactivity with benzo[b]fluoranthene (2.7 %), other interference to the assay was negligible ( 0.05 %). The validity of BaP kit in 4 A degrees C was above 6 months. The sensitivity, specificity, simplicity, and reliability of BaP kit were confirmed by the procedure utilizing gas chromatography coupled to mass spectrometric (GC-MS) for the identification and quantification of BaP in barbecue. Their coincidence rate was 100 % compared with GC-MS. The kit was proved to be used for the rapid determination of BaP residues.
机译：本文介绍了通过细胞融合程序生成对苯并py（BaP）具有高特异性的单克隆抗体（mAb），以及使用这些杂交瘤之一开发基于mAb的间接竞争性酶联免疫吸附试剂盒（icELISA试剂盒）的方法（克隆2E12）。含标准BaP抑制剂的BaP-Kit的校准曲线为典型的S型曲线，符合四个参数对数方程，线性回归方程y = -28.662x + 132.87，半最大抑制浓度（IC50）为0.779μg / L灵敏度为0.054μg/ L。牛肉中添加的BaP的回收率在81.3％至93.6％之间。通过批间和批内系数变化确定的测定精度和准确性低于6％。除了与苯并[b]荧蒽的轻微交叉反应（2.7％）外，对测定的其他干扰可忽略不计（<0.05％）。 BaP试剂盒在4 A摄氏度下的有效性超过6个月。 BaP试剂盒的灵敏度，特异性，简便性和可靠性已通过使用气相色谱-质谱联用（GC-MS）进行烧烤中BaP鉴定和定量的程序进行了确认。与GC-MS相比，它们的符合率为100％。证明该试剂盒可用于快速测定BaP残留量。

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