Sullivan T J, Hechemy K E, Harris H L, Rudofsky U H, Samsonoff W A, Peterson A J, Evans B D, Balaban S L
David Axelrod Institute for Public Health/Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
J Clin Microbiol. 1994 Feb;32(2):423-9. doi: 10.1128/jcm.32.2.423-429.1994.
We have produced, by using a sonicate of Borrelia burgdorferi, a monoclonal antibody (MAb), NYSP39H, that is specific for the P39 protein band. This MAb reacted with 13 isolates of B. burgdorferi but not with eight different spirochetes (four borrelias, two leptospiras, and two treponemas). Surface labeling of B. burgdorferi with biotin and subsequent treatment with Nonidet P-40 showed that P39 was not biotinylated but was extracted with Nonidet P-40, indicating that it is present within the outer membrane, but not on the surface of the spirochete. Immunoelectron microscopy revealed the immunogold probe primarily at the cytoplasmic membrane region of the spirochete. The MAb detected B. burgdorferi in the indirect fluorescent-antibody test only when the spirochetes from a culture or in a tick homogenate were fixed with polylysine and not with acetone. NYSP39H appears to be an appropriate probe for use in the specific detection of B. burgdorferi.
我们通过使用伯氏疏螺旋体的超声裂解物制备了一种单克隆抗体（MAb）NYSP39H，它对P39蛋白条带具有特异性。该单克隆抗体与13株伯氏疏螺旋体发生反应，但不与8种不同的螺旋体（4种疏螺旋体、2种钩端螺旋体和2种密螺旋体）发生反应。用生物素对伯氏疏螺旋体进行表面标记，随后用去氧胆酸钠处理，结果表明P39未被生物素化，但可被去氧胆酸钠提取，这表明它存在于外膜内，而不在螺旋体表面。免疫电子显微镜显示免疫金探针主要位于螺旋体的细胞质膜区域。仅当来自培养物或蜱匀浆中的螺旋体用聚赖氨酸而非丙酮固定时，该单克隆抗体才能在间接荧光抗体试验中检测到伯氏疏螺旋体。NYSP39H似乎是用于特异性检测伯氏疏螺旋体的合适探针。
