Fang Rong, Houhamdi Linda, Raoult Didier
Unité des Rickettsies, CNRS UMR 6020, IFR 48, Faculty of Medicine, 13385 Marseille, France.
J Clin Microbiol. 2002 Sep;40(9):3358-63. doi: 10.1128/JCM.40.9.3358-3363.2002.
In order to identify Rickettsia prowazekii in lice, we developed a panel of 29 representative monoclonal antibodies selected from 187 positive hybridomas made by fusing splenocytes of immunized mice with SP2/0-Ag14 myeloma cells. Immunoblotting revealed that 15 monoclonal antibodies reacted with the lipopolysaccharide-like (LPS-L) antigen and 14 reacted with the epitopes of a 120-kDa protein. Only typhus group rickettsiae reacted with the monoclonal antibodies against LPS-L. R. felis, a recently identified rickettsial species, did not react with these monoclonal antibodies, confirming that it is not antigenically related to the typhus group. Monoclonal antibodies against the 120-kDa protein were highly specific for R. prowazekii. We successfully applied a selected monoclonal antibody against the 120-kDa protein to detect by immunofluorescence assay R. prowazekii in smears from 56 wild and laboratory lice, as well as in 10 samples of louse feces infected or not infected with the organism. We have developed a simple, practical, and specific diagnostic assay for clinical specimens and large-scale epidemiological surveys with a sensitivity of 91%. These monoclonal antibodies could be added to the rickettsial diagnostic panel and be used to differentiate R. prowazekii from other rickettsial species.
为了在虱子中鉴定普氏立克次体,我们从187个阳性杂交瘤中筛选出一组29种具有代表性的单克隆抗体,这些杂交瘤是通过将免疫小鼠的脾细胞与SP2/0-Ag14骨髓瘤细胞融合制备而成的。免疫印迹显示,15种单克隆抗体与脂多糖样(LPS-L)抗原发生反应,14种与一种120 kDa蛋白的表位发生反应。只有斑疹伤寒群立克次体能与抗LPS-L的单克隆抗体发生反应。最近鉴定出的立克次体物种——猫立克次体,不与这些单克隆抗体发生反应,这证实了它在抗原性上与斑疹伤寒群无关。抗120 kDa蛋白的单克隆抗体对普氏立克次体具有高度特异性。我们成功地应用一种针对120 kDa蛋白的单克隆抗体,通过免疫荧光试验在56只野生和实验室虱子的涂片以及10份感染或未感染该病原体的虱子粪便样本中检测到了普氏立克次体。我们已经开发出一种简单、实用且特异的诊断方法,用于临床标本和大规模流行病学调查,灵敏度为91%。这些单克隆抗体可添加到立克次体诊断试剂组中,用于区分普氏立克次体与其他立克次体物种。