抗微小螺旋体（ATCC 29335）多克隆抗血清的制备与鉴定

Production and characterization of a polyclonal antiserum against Spiroplasma mirum (ATCC 29335).

作者信息

Tenckhoff B, Kölmel H W, Wolf V, Lange R

机构信息

Immanuel-Krankenhaus GmbH, Rheumaklinik, Berlin (Wannsee), Germany.

出版信息

Zentralbl Bakteriol. 1994 Jan;280(3):409-15. doi: 10.1016/s0934-8840(11)80605-5.

DOI:10.1016/s0934-8840(11)80605-5

PMID:8167436

Abstract

Spiroplasma mirum (ATCC 29335), an American tick isolate, was cultivated and its purity was controlled using electron microscopy and Tricine-SDS-PAGE. In the SDS-PAGE analysis, the protein pattern of the antigen used revealed a polypeptide profile (35 bands) with an approximate molecular weight of 161 kDa to 10.5 kDa. This strain was used to immunize two rabbits. The produced antisera did not cross-react with Mycoplasma pneumoniae, Salmonella minnesota, Borrelia burgdorferi, Treponema pallidum but did still react in Western blot analysis with Spiroplasma mirum at a dilution of 1:128.000. This specific and sensitive antiserum was used to examine 72 midgut smears of Berlin ticks (lxodes ricinus) individually by direct immunofluorescence. In 26 samples, we could demonstrate fluorescent structures, but we were not able to culture spiroplasmas from these samples.

摘要

微小螺旋体（ATCC 29335），一种从美国蜱虫中分离出的菌株，进行了培养，并通过电子显微镜和Tricine-SDS-PAGE控制其纯度。在SDS-PAGE分析中，所用抗原的蛋白质图谱显示出一条多肽谱（35条带），分子量约为161 kDa至10.5 kDa。该菌株用于免疫两只兔子。所产生的抗血清与肺炎支原体、明尼苏达沙门氏菌、伯氏疏螺旋体、梅毒螺旋体无交叉反应，但在Western blot分析中，在1:128,000的稀释度下仍与微小螺旋体发生反应。这种特异性和敏感性的抗血清用于通过直接免疫荧光分别检测72份柏林蜱（蓖麻硬蜱）中肠涂片。在26个样本中，我们能够证明存在荧光结构，但无法从这些样本中培养出螺旋体。