Chen Fur-Chi, Hsieh Y H Peggy, Bridgman Roger C
Institute of Agricultural and Environmental Research, Tennessee State University, 3500 John A. Merritt Boulevard, Nashville, Tennessee 37209, USA.
J Food Prot. 2004 Mar;67(3):544-9. doi: 10.4315/0362-028x-67.3.544.
Regulations aimed to control the epidemic of bovine spongiform encephalopathy have banned the use of certain animal products, i.e., ruminant meat and bone meals, in ruminant animal feeds. A sensitive enzyme-linked immunosorbent assay has been developed to detect prohibited bovine and ovine muscles in feedstuffs. The assay utilizes a pair of monoclonal antibodies (MAbs) against skeletal troponin I (TnI). MAb 5G9, specific to bovine and ovine TnI, was used as the capture antibody and the biotin-conjugated MAb 2G3, reacting to all heterologous TnI, was used as the detection antibody. Quantitative procedures were applied to samples containing 5, 0.5, and 0.05% (wt/wt) of heat-treated (132 degrees C/2 bar, 2 h) bovine and ovine meat meals in three different feeds, coexisting with porcine, chicken, or turkey meat meal. The presence of these nonprohibited species did not affect the detection of bovine and ovine meat meals in the feed samples (P > 0.05). Quantitative determinations of extractable bovine and ovine TnI, with a detection limit of 5.0 and 4.0 ng/ml, respectively, were achieved when the matching feed matrixes were used in the calibration curves. This new assay provides a rapid and reliable way to detect animal protein products containing a trace amount of bovine or ovine muscle tissue in feedstuffs.
旨在控制牛海绵状脑病疫情的法规已禁止在反刍动物饲料中使用某些动物产品，即反刍动物肉骨粉。已开发出一种灵敏的酶联免疫吸附测定法来检测饲料中违禁的牛肉和羊肉。该测定法利用一对针对骨骼肌肌钙蛋白I（TnI）的单克隆抗体（MAb）。对牛和羊TnI具有特异性的单克隆抗体5G9用作捕获抗体，与所有异源TnI反应的生物素偶联单克隆抗体2G3用作检测抗体。对三种不同饲料中含有5%、0.5%和0.05%（重量/重量）热处理（132摄氏度/2巴，2小时）牛肉和羊肉粉的样品进行了定量分析，这些样品与猪肉、鸡肉或火鸡肉粉共存。这些非违禁物种的存在不影响饲料样品中牛肉和羊肉粉的检测（P>0.05）。当在校准曲线中使用匹配的饲料基质时，分别实现了可提取的牛和羊TnI的定量测定，检测限分别为5.0和4.0纳克/毫升。这种新的测定法提供了一种快速可靠的方法来检测饲料中含有微量牛或羊肌肉组织的动物蛋白产品。
