Murata M, Yoshida Y, Osono M, Ohashi N, Oyanagi M, Urakami H, Tamura A, Nogami S, Tanaka H, Kawamura A
Microbiol Immunol. 1986;30(7):599-610. doi: 10.1111/j.1348-0421.1986.tb02987.x.
We have developed 18 hybridoma cell lines which secrete murine monoclonal strain-specific antibodies to prototype strains of Rickettsia tsutsugamushi: nine anti-Gilliam, four anti-Karp and five anti-Kato antibodies. All the monoclonal antibodies reacted only with their homologous strains in direct and indirect immunofluorescence (IF), or indirect immunoperoxidase (IP) test. By IF and IP tests with the monoclonal antibodies, 22 strains of R. tsutsugamushi, which were newly isolated from mites, field rodents and patients with Tsutsugamushi disease (scrub typhus) in Japan, were all clearly identified as either Gilliam or Karp type. Analysis by polyacrylamide gel electrophoresis and immunoblotting techniques revealed that the monoclonal antibodies recognized primarily the polypeptides of an apparent molecular weight of 54 to 56 kilodaltons of the homologous rickettsial surface. The monoclonal antibodies produced in the present study should enhance the serotyping and further analytical investigation of the rickettsial antigens since they recognize the strain- or type-specific polypeptides and do not show any cross-reaction among strains.
我们已经开发出18种杂交瘤细胞系,它们能分泌针对恙虫病立克次体原型菌株的鼠单克隆菌株特异性抗体:9种抗吉列姆抗体、4种抗卡尔普抗体和5种抗加藤抗体。在直接和间接免疫荧光(IF)或间接免疫过氧化物酶(IP)试验中,所有单克隆抗体仅与其同源菌株发生反应。通过使用单克隆抗体进行IF和IP试验,从日本的螨虫、野外啮齿动物和恙虫病(丛林斑疹伤寒)患者中新分离出的22株恙虫病立克次体菌株,均被明确鉴定为吉列姆型或卡尔普型。聚丙烯酰胺凝胶电泳和免疫印迹技术分析表明,单克隆抗体主要识别同源立克次体表面表观分子量为54至56千道尔顿的多肽。本研究中产生的单克隆抗体应能加强对立克次体抗原的血清分型和进一步的分析研究,因为它们识别菌株或型特异性多肽,且在菌株之间不显示任何交叉反应。