Ruby K W, Cardella M A, Knudtson W U
National Veterinary Services Laboratories, U.S. Department of Agriculture, Ames, Iowa 50010.
Biologicals. 1992 Dec;20(4):259-66. doi: 10.1016/s1045-1056(05)80045-4.
An enzyme-linked immunosorbent assay (ELISA) was developed for the quantitation of leptospiral antigen in bacterins containing Leptospira interrogans serovar pomona type kennewicki. A monoclonal antibody (MAb), 2D7, which is directed against a surface antigen on whole cells of L. interrogans serovar pomona type kennewicki, was used in the assay. The capture of antigen in bacterins by a polyclonal antiserum was followed by the addition of the 2D7 ascites fluid, an anti-mouse conjugate and substrate. Biologicals evaluated with this system included preparations containing type kennewicki antigen (homologous) and those not containing type kennewicki antigen (heterologous). Heterologous bacterins gave optical density (OD) values comparable to those of blank wells. Homologous bacterins yielded OD values equal to or greater than those of the National Veterinary Services Laboratories (NVSL) reference pomona bacterin. The relative potencies (RP) of 84 licensed commercial Leptospira pomona bacterin serials were evaluated against the NVSL reference pomona bacterin using the NVSL Relative Potency computer program. Random samples of 1, 2, 3 and 5 ml dose products were selected for evaluation with this system. All products tested passed the hamster potency assay required for leptospiral bacterins. This ELISA system enables detection of antigen in bacterins containing L. interrogans serovar pomona type kennewicki and demonstrates the potential for in vitro testing of leptospiral bacterins.
开发了一种酶联免疫吸附测定法（ELISA），用于定量检测含有波摩那钩端螺旋体肯尼维奇型的菌苗中的钩端螺旋体抗原。该测定法使用了一种单克隆抗体（MAb）2D7，它针对波摩那钩端螺旋体肯尼维奇型全细胞表面抗原。先用多克隆抗血清捕获菌苗中的抗原，然后加入2D7腹水、抗小鼠缀合物和底物。用该系统评估的生物制品包括含有肯尼维奇型抗原的制剂（同源）和不含肯尼维奇型抗原的制剂（异源）。异源菌苗的光密度（OD）值与空白孔相当。同源菌苗产生的OD值等于或大于国家兽医服务实验室（NVSL）的参考波摩那菌苗。使用NVSL相对效力计算机程序，针对NVSL参考波摩那菌苗评估了84个已获许可的商业波摩那钩端螺旋体菌苗系列的相对效力（RP）。选择1、2、3和5毫升剂量产品的随机样本用该系统进行评估。所有测试产品均通过了钩端螺旋体菌苗所需的仓鼠效力试验。该ELISA系统能够检测含有波摩那钩端螺旋体肯尼维奇型的菌苗中的抗原，并证明了钩端螺旋体菌苗体外检测的潜力。
