Belzer C A, Tabatabai L B, Deyoe B L
U.S. Department of Agriculture, National Animal Disease Center, Ames, IA 50010.
Vet Microbiol. 1991 Mar;27(1):79-90. doi: 10.1016/0378-1135(91)90064-m.
Brucella abortus strain 19 salt-extractable proteins fractionated by differential ammonium sulfate precipitation were used in a western blotting method to detect bovine immunoglobulin G antibodies to B. abortus. Sera from infected cattle and from cattle vaccinated with strain 19 and subsequently exposed to virulent B. abortus bound to a common group of antigens ranging in molecular weights from 31,000 to 45,000 daltons. Immunoglobulin G antibodies in sera from the latter group in addition also bound to antigens with molecular weights of 66,000 to 71,000 daltons. Some sera from cattle vaccinated when sexually mature reacted similar to those from infected cattle, while immunoglobulin G antibodies in sera from Brucella-free cattle and vaccinated calves did not bind to either group of antigens. In general, fractionation of the proteins by ammonium sulfate precipitation offered no advantage for detecting differences between groups of sera. Ammonium sulfate fraction 0 to 35% reacted with a larger number of sera from a naturally infected group than fraction 0 to 70%. Both fractions reacted equally well with sera from the other groups of cattle, while fractions 35 to 70% and 70 to 100% reacted poorly in this technique. The attractive feature of the blot is that sera from calfhood-vaccinated cattle did not react.
用硫酸铵分级沉淀法分离出的流产布鲁氏菌19株盐提取蛋白，采用蛋白质印迹法检测牛抗流产布鲁氏菌免疫球蛋白G抗体。感染牛的血清以及接种19株菌并随后接触强毒流产布鲁氏菌的牛的血清，与一组分子量在31,000至45,000道尔顿之间的共同抗原结合。后一组牛血清中的免疫球蛋白G抗体还与分子量为66,000至71,000道尔顿的抗原结合。一些性成熟时接种疫苗的牛的血清反应与感染牛的血清相似，而未感染布鲁氏菌的牛和接种疫苗的小牛血清中的免疫球蛋白G抗体均不与这两组抗原结合。一般来说，硫酸铵沉淀法分离蛋白质对于检测不同组血清之间的差异并无优势。硫酸铵分级沉淀0至35%部分与自然感染组的更多血清发生反应，而0至70%部分则不然。这两个部分与其他组牛的血清反应同样良好，而35至70%和70至100%部分在此技术中反应较差。该印迹法的吸引人之处在于，幼年接种疫苗的牛的血清不发生反应。
