Rapid quantification of melamine in milk using competitive 1,1′-oxalyldiimidazole chemiluminescent enzyme immunoassay
Rapid quantification of melamine in milk using competitive 1,1′-oxalyldiimidazole chemiluminescent enzyme immunoassay
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DOI：
10.1039/c0an00396d
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发表时间：
2010-01-01
期刊：
影响因子：
4.2
通讯作者：
Lee, Ji Hoon
中科院分区：
文献类型：
--
作者：
Choi, JooHee;Kim, Young-Teck;Lee, Ji Hoon
建立了一种新的竞争性1，1 '-草酰二咪唑（ODI）荧光酶免疫分析法（CLEIA），用于牛奶中三聚氰胺的快速检测。在三聚氰胺结合辣根过氧化物酶的存在下，牛奶中存在的脂肪在三聚氰胺和抗三聚氰胺的竞争性结合相互作用中起抑制剂的作用。因此，竞争ODI CLEIA的校准曲线和灵敏度的定量三聚氰胺在无脂牛奶比那些在含脂肪的牛奶更广泛和更好。然而，离心不是去除抑制剂的好方法，因为一部分三聚氰胺也会随着脂肪一起去除。在pH 7.4的PBS缓冲液稀释的20%牛奶中，抗-三聚氰胺与三聚氰胺竞争结合作用的孵育时间（20 min）比在100%牛奶中的孵育时间（10 min）长，但前者的灵敏度比后者高。在快速ODI CLEIA（动态范围：3.8-125 ppb）中确定的检测限（1.12 ppb）的定量三聚氰胺在20%的牛奶不含脂肪低于那些（6.3和9.0 ppb）计算相对耗时的鲁米诺CLEIA和酶联免疫吸附试验（ELISA）。此外，我们期望ODI-CLEIA（动态范围：62.5-2000 ppb）能够直接定量100%牛奶中的三聚氰胺，而无需任何预处理，可以作为一种新的和简单的方法，用于快速筛查牛奶中的三聚氰胺。
A novel competitive 1,1'-oxalyldiimidazole (ODI) chemiluminescent enzyme immunoassay (CLEIA) was developed as a method for rapid and simple screening of melamine in milk. Fat existing in milk acts as an inhibitor in the competitive binding interaction of melamine and anti-melamine in the presence of melamine-conjugated horseradish peroxidase. Thus, the calibration curve and sensitivity of competitive ODI CLEIA for the quantification of melamine in fat free milk were wider and better than those in milk containing fat. However, a centrifuge is not a good method for removing the inhibitor because a portion of the melamine is also removed with the fat. The incubation time (20 min) for the competitive binding interaction of anti-melamine and melamine in 20% milk diluted with PBS buffer of pH 7.4 was longer than that (10 min) in 100% milk even though the sensitivity of the former was better than latter. The limit of detection (1.12 ppb) determined in rapid ODI CLEIA (dynamic range: 3.8-125 ppb) for the quantification of melamine in 20% milk not containing fat was lower than those (6.3 and 9.0 ppb) calculated in relatively time-consuming luminol CLEIA and enzyme-linked immunosorbent assay (ELISA). Also, we expect that ODI-CLEIA (dynamic range: 62.5-2000 ppb) capable of directly quantifying melamine in 100% milk without any pretreatment can be applied as a new and simple method for rapid screening of melamine in milk.