Raoult D, Dasch G A
Centre National de Reference des Rickettsioses, C.H.U. La Timone, Marseille, France.
J Immunol Methods. 1989 Dec 20;125(1-2):57-65. doi: 10.1016/0022-1759(89)90078-1.
A procedure is described for assaying antibodies based on the application of antigen to nitrocellulose as a line with an ink pen point. The method requires no expensive apparatus, is easy to perform, and requires less than 0.25 micrograms of antigen per assay. More than 45 antigens can be assayed simultaneously with a single antibody. Antigens can be applied as purified proteins, extracts, or sodium dodecyl sulfate solubilized extracts. The application of the line blot assay for the detection of monoclonal antibodies which recognize heat-sensitive and insensitive epitopes on the typhus rickettsia surface protein antigen is described. A new serotyping assay for Gram-negative bacteria is also described in which sodium dodecyl sulfate solubilized antigens are applied as lines with and without prior proteinase K digestion. The value of the line blot serotyping assay is demonstrated with Proteus. Rickettsia, Rochalimaea, and Legionella antigens. The line blot immunoassay is a simple, but powerful and flexible, alternative to dot and cross-dot immunoassays.
本文描述了一种基于用墨水笔尖将抗原划成线涂覆在硝酸纤维素膜上来检测抗体的方法。该方法不需要昂贵的仪器，操作简便，每次检测所需抗原量少于0.25微克。一种抗体可同时检测45种以上的抗原。抗原可以是纯化蛋白、提取物或十二烷基硫酸钠溶解的提取物。文中描述了线印迹分析用于检测识别斑疹伤寒立克次体表面蛋白抗原上热敏感和热不敏感表位的单克隆抗体。还介绍了一种针对革兰氏阴性菌的新型血清分型检测方法，其中十二烷基硫酸钠溶解的抗原在蛋白酶K消化前后划成线涂覆。用变形杆菌、立克次体、罗卡利马氏体和军团菌抗原证明了线印迹血清分型检测的价值。线印迹免疫分析是斑点免疫分析和交叉斑点免疫分析的一种简单、但强大且灵活的替代方法。
