Hulínská D, Krausová M, Janovská D, Rohácová H, Hancil J, Mailer H
Department of Electron Microscopy, National Institute of Public Health, Prague, Czech Republic.
Cent Eur J Public Health. 1993 Dec;1(2):81-5.
Results of studies using direct antigen detection suggest that seronegative Lyme borreliosis is not rare and support the hypothesis that Borrelia antigens can persist in humans. We report three successful cultures from blood out of 30 attempts from 96 Lyme disease patients. The proof of borreliaemia in early or late phases of Lyme disease by immuno-capture electron microscopy has practical importance for subsequent cultivation. The polymerase chain reaction with oligonucleotide sequences directed against 16S rRNA identified two of our blood isolates as Borrelia burgdorferi genospecies III., VS 461 group, and one as Borrelia garinii sp. nov. All of the three isolates were reactive with monoclonal antibody H9724 against flagellin and with antibody against main extracellular protein at 83 kDa. Borrelia garinii had a single predominant protein OspA at 33.5 kDa and reacted with monoclonal antibody H5332 in contrast to two isolates of the VS 461 group with two major proteins OspA and OspB at 32.5 and 35 kDa. We conclude that isolation of spirochetes from the blood might prove successful in clinically selected cases of Lyme borreliosis. Immuno-capture electron microscopy has proved to be a sensitive assay for monitoring and studying Lyme borreliosis.
使用直接抗原检测的研究结果表明，血清阴性莱姆病并不罕见，并支持伯氏疏螺旋体抗原可在人体内持续存在的假说。我们报告了在对96例莱姆病患者进行的30次血液培养尝试中，有3次成功培养出病原体。通过免疫捕获电子显微镜在莱姆病早期或晚期证实存在菌血症，对后续培养具有实际意义。针对16S rRNA的寡核苷酸序列进行的聚合酶链反应鉴定出我们的两份血液分离株为伯氏疏螺旋体基因种III.，VS 461组，另一份为新种加氏疏螺旋体。所有这三份分离株都与抗鞭毛蛋白的单克隆抗体H9724以及抗83 kDa主要细胞外蛋白的抗体发生反应。加氏疏螺旋体有一个单一的主要外膜蛋白OspA，分子量为33.5 kDa，并与单克隆抗体H5332发生反应，而VS 461组的两份分离株有两种主要外膜蛋白OspA和OspB，分子量分别为32.5 kDa和35 kDa。我们得出结论，在临床选择的莱姆病病例中，从血液中分离螺旋体可能会成功。免疫捕获电子显微镜已被证明是监测和研究莱姆病的一种灵敏检测方法。
