Development of a Specifically Enhanced Enzyme-Linked Immunosorbent Assay for the Detection of Melamine in Milk
Development of a Specifically Enhanced Enzyme-Linked Immunosorbent Assay for the Detection of Melamine in Milk
复制标题
开发用于检测牛奶中三聚氰胺的特异性增强酶联免疫吸附测定法
DOI：
10.3390/molecules16075591
复制
发表时间：
2011-06-30
期刊：
影响因子：
4.6
通讯作者：
Sun Y
中科院分区：
文献类型：
--
作者：
Lei H;Su R;Haughey SA;Wang Q;Xu Z;Yang J;Shen Y;Wang H;Jiang Y;Sun Y
建立了一种检测牛奶中三聚氰胺的间接竞争酶联免疫吸附试验(IcEL ISA)。用三种不同间隔臂的三聚氰胺半抗原制备不同的平板包被抗原。结果表明，以3-(4，6-diamino-1，6-dihydro-1，3，5-triazin-2-ylthio)propanoic酸(半抗原C)与卵清蛋白(OVA)偶联制备的包被抗原对三聚氰胺具有最好的敏感性和特异性。其半数抑制浓度(IC50值)为35.4 ng·m L−1，检出限为8.9 ng·m L−1，检测范围(20~80%抑制浓度)为14.9~108.5 ng·m L−1。与先前报道的酶联免疫吸附试验结果相比，本研究中开发的间接酶联免疫吸附试验对环丙嗪的交叉反应性要低得多，环丙嗪是一种广泛用于蔬菜和马厩的灭蝇杀虫剂。从牛奶样品中获得的回收率与用高效液相-质谱仪测定的结果一致，表明该方法的检测性能能够满足食品法典委员会制定的残留限量的要求。因此，建立的免疫分析方法可用于牛奶中三聚氰胺的分析。
An indirect competitive enzyme-linked immunosorbent assay (icELISA) with enhanced specificity for melamine in milk was developed. Three haptens of melamine with different spacer-arms were used to prepare different plate coating antigens. It was found that the icELISA show best sensitivity and specificity to melamine when using the coating antigen prepared by coupling 3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylthio)propanoic acid (Hapten C) with ovalbumin (OVA). The 50% inhibitory concentration (IC50) value was 35.4 ng·mL−1, the limit of detection (LOD) was 8.9 ng·mL−1 and the detectable working range (20–80% inhibitory concentration) was from 14.9 to 108.5 ng·mL−1, respectively. Compared to the ELISA results previously reported, the developed icELISA in the present study showed a much lower cross-reactivity to cyromazine, a fly-killing insecticide widely used in vegetables and stables. Recoveries obtained from milk samples in this study were in agreement with those obtained using the HPLC-MS method, indicating the detection performance of the icELISA could meet the requirement of the residue limit set by the Codex Alimentarius Commission. Therefore, the developed immunoassay can be applied for the analysis of melamine presented in milk.
登录
查看更多内容
影响因子：
3
作者：
Kato, Mihoko;Ihara, Yoshiharu;Nakazawa, Hiroyuki
通讯作者：
Nakazawa, Hiroyuki
影响因子：
6.1
作者：
Zhu, Xinle;Wang, Shuhuai;Chen, Huilan
通讯作者：
Chen, Huilan
影响因子：
4.2
作者：
Choi, JooHee;Kim, Young-Teck;Lee, Ji Hoon
通讯作者：
Lee, Ji Hoon
DOI：
10.1080/19440040903289720
发表时间：
2010-01-01
影响因子：
2.9
作者：
Tittlemier, S. A.
通讯作者：
Tittlemier, S. A.
DOI：
10.2174/138620703106298310
发表时间：
2003-05-01
影响因子：
1.8
作者：
Nasir, MS;Jolley, ME
通讯作者：
Jolley, ME