Animal Parasitic Diseases Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, USDA, Beltsville, MD 20705, USA.
Res Vet Sci. 2011 Dec;91(3):e80-6. doi: 10.1016/j.rvsc.2011.01.017. Epub 2011 Feb 22.
Clostridium-related poultry diseases such as necrotic enteritis (NE) and gangrenous dermatitis (GD) cause substantial economic losses on a global scale. Two antigenic Clostridium perfringens proteins, elongation factor Tu (EF-Tu) and pyruvate:ferredoxin oxidoreductase (PFO), were identified by reaction with immune sera from commercial meat-type chickens with clinical outbreak of Clostridium infections. In addition to the genes encoding EF-Tu and PFO, C. perfringens alpha-toxin and necrotic enteritis B-like (NetB) toxin were also expressed in Escherichia coli and their corresponding recombinant proteins were purified. Using the four recombinant proteins as target antigens in ELISA immunoassays, high serum antibody titers were observed not only in chickens with clinical signs of Clostridium infections, but also in apparently healthy animals from the same disease-endemic farm. By contrast, no antibodies against any of the proteins were present in the serum of a specific pathogen-free bird. In ELISA using recombinant proteins of C. perfringens, the levels of anti-bacterial protein antibodies were also higher in chickens which were experimentally induced to show NE clinical signs after co-infection with C. perfringens and Eimeria maxima compared with uninfected controls. These results show that two antigenic C. perfringens proteins, EF-Tu and PFO can be useful detection antigens for C. perfringens-afflicted infections in commercial poultry.
与梭菌相关的家禽疾病，如坏死性肠炎（NE）和坏疽性皮炎（GD），在全球范围内造成了巨大的经济损失。通过与患有梭菌感染临床症状的商业肉用鸡的免疫血清反应，鉴定出两种抗原性梭状芽孢杆菌蛋白，延伸因子 Tu（EF-Tu）和丙酮酸：铁氧还蛋白氧化还原酶（PFO）。除了编码 EF-Tu 和 PFO 的基因外，梭状芽孢杆菌α-毒素和坏死性肠炎 B 样（NetB）毒素也在大肠杆菌中表达，并纯化了相应的重组蛋白。使用这四种重组蛋白作为 ELISA 免疫分析的靶抗原，不仅在患有梭菌感染临床症状的鸡中，而且在同一疾病流行农场的明显健康动物中，都观察到了高血清抗体滴度。相比之下，在无特定病原体鸟类的血清中，不存在针对任何一种蛋白质的抗体。在使用梭状芽孢杆菌重组蛋白的 ELISA 中，在与巨型艾美耳球虫共感染后表现出 NE 临床症状的鸡中，针对细菌蛋白的抗体水平也高于未感染对照鸡。这些结果表明，两种抗原性梭状芽孢杆菌蛋白 EF-Tu 和 PFO 可作为商业家禽梭状芽孢杆菌感染的有用检测抗原。
