Department of Chemistry and Pharmacy, Food Chemistry, Emil Fischer Center, University of Erlangen-Nuremberg, Schuhstrasse 19, 91052 Erlangen, Germany.
J Agric Food Chem. 2010 Jun 9;58(11):6587-93. doi: 10.1021/jf100625g.
Bovine spongiform encephalopathy (BSE) is transmitted by the ingestion of central nervous system (CNS) tissue of infected animals. Food inspection must, therefore, test for the presence of CNS tissue in meat and meat products. A Western blot assay for the specific CNS tissue marker myelin proteolipid protein (PLP) was optimized with considerably reduced analysis time, solvent consumption, and detection limit (0.001% CNS tissue in minced beef). Further, a PLP-specific recombinant bivalent fragment antigen binding mini-antibody (anti-PLP Fab) was obtained from a commercial phage display library. Western blot analysis with the anti-PLP Fab selectively detected CNS tissue in minced beef with a detection limit of 0.025%. Model experiments for meat processing revealed that assay sensitivity decreased with increasing temperature and prolonged heating time. A market survey with 687 sausage samples was performed using PLP-Western blot and enzyme-linked immunosorbent assay (ELISA) for glial fibrillary acidic protein (GFAP). Five samples were tested clearly positive by both assay systems, whereas in an additional six samples, CNS tissue was detected only by GFAP ELISA and in two samples only by PLP-Western blot.
牛海绵状脑病（BSE）通过摄入感染动物的中枢神经系统（CNS）组织传播。因此，食品检验必须检测肉类和肉类产品中是否存在中枢神经系统组织。一种针对中枢神经系统组织标志物髓鞘少突胶质细胞糖蛋白（PLP）的 Western blot 检测方法经过优化，分析时间、溶剂消耗和检测限（切碎牛肉中 0.001%的中枢神经系统组织）均显著降低。此外，还从商业噬菌体展示文库中获得了针对 PLP 的重组二价片段抗原结合 mini 抗体（抗-PLP Fab）。用抗-PLP Fab 进行的 Western blot 分析选择性地检测到切碎牛肉中的中枢神经系统组织，检测限为 0.025%。肉类加工的模型实验表明，随着温度的升高和加热时间的延长，检测灵敏度会降低。使用 PLP-Western blot 和酶联免疫吸附试验（ELISA）检测神经胶质纤维酸性蛋白（GFAP）对 687 个香肠样品进行了市场调查。两种检测系统均检测到 5 个样本呈明显阳性，而在另外 6 个样本中，仅 GFAP ELISA 检测到中枢神经系统组织，在 2 个样本中仅通过 PLP-Western blot 检测到。
