胭脂红酸ELISA检测试剂盒的制备及应用

doi:10.16389/j.cnki.cn42-1737/n.2019.05.008

江汉大学学报（自然科学版） ›› 2019, Vol. 47 ›› Issue (5): 436-441.doi: 10.16389/j.cnki.cn42-1737/n.2019.05.008

胭脂红酸ELISA检测试剂盒的制备及应用

李佳楠¹，杨帆帆²，何峰容^*1，2

1. 江汉大学生命科学学院，湖北武汉 430056；2. 武汉云克隆科技股份有限公司，湖北武汉 430056

发布日期:2019-10-08
通讯作者: 何峰容
作者简介:李佳楠（1978—），女，教授，博士，研究方向：生物技术。
基金资助:
武汉市黄鹤英才计划资助项目（2014）

Preparation and Application of Carminic Acid ELISA Kit

LI Jianan¹，YANG Fanfan²，HE Fengrong^*1，2

1. School of Life Sciences，Jianghan University，Wuhan 430056，Hubei，China；2. Wuhan Cloud-Clone Co. ，Ltd. ，Wuhan 430056，Hubei，China

Published:2019-10-08
Contact: HE Fengrong

摘要/Abstract

摘要： 为了快速高效检测肉类食品中胭脂红酸含量，制备了胭脂红酸ELISA检测试剂盒，检测了市售肉类样品中胭脂红酸含量，并通过高效液相色谱法进行了验证。结果显示试剂盒的检测限为1 ng/mL，线性范围7. 8 ~ 1 150 ng/mL，灵敏度IC₅₀值为95. 2 ng/mL；该试剂盒中的检测抗体对胭脂红的交叉反应率为108. 7%，但与其他4 种色素添加剂均未见交叉反应。检测市售肉类样品的批内回收率在79. 1% ~ 103. 1% 之间，批间回收率在81. 6% ~ 98. 8%之间；批内、批间试验的相对标准偏差基本小于10%，且与HPLC 方法具有良好的一致性（R² =0. 927），说明该试剂盒具有灵敏、准确、简便、快速的特点，为胭脂红酸免疫学快速检测方法的建立提供了技术基础。

关键词: 胭脂红酸, 酶联免疫吸附试验（ELISA）, 多克隆抗体, 高效液相色谱（HPLC）

Abstract: In order to detect carminic acid（CA）content in meat product with high sensitivity and specificity，an ELISA kit for CA was prepared to detect the CA content in meat product of market sale. At the same time，the detection situation was further confirmed by high- performance liquid chromatography（HPLC）. The results showed that the dynamic range was from 7. 8 ~ 1 150 ng/mL with the limit of detection（LOD）1 ng/mL，and 50% inhibitive concentration（IC₅₀）of CA was 95. 2 ng/mL；the cross-reactivity of the PAB toward carmine was 108. 7%，and no response was found with other four edible colorants. The spiked meat samples from meat product of market sale were detected by ELISA with intra- and inter-assay recovery ranges of 79. 1% ~ 103. 1% and 81. 6% ~ 98. 8% ，respectively. The relative standard deviations of intra- and inter-assay were less than 10%. Finally，the results for the samples were further confirmed by high- performance liquid chromatography（HPLC）with a high correlation coefficient of 0. 927，these demonstrated that the method in the study was sensitive， accurate， simple and fast， the method established the technical foundation for immunological rapid detection of carminic acid.

Key words: carminic acid, enzyme linked immunosorbent assay（ELISA）, polyclonal antibody, highperformance liquid chromatography（HPLC）

中图分类号:

R155.51

李佳楠，杨帆帆，何峰容. 胭脂红酸ELISA检测试剂盒的制备及应用[J]. 江汉大学学报（自然科学版）, 2019, 47(5): 436-441.

LI Jianan，YANG Fanfan，HE Fengrong. Preparation and Application of Carminic Acid ELISA Kit[J]. Journal of Jianghan University (Natural Science Edition), 2019, 47(5): 436-441.

胭脂红酸ELISA检测试剂盒的制备及应用

Preparation and Application of Carminic Acid ELISA Kit

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