Halbgebauer Steffen, Haußmann Ute, Klafki Hans, Tumani Hayrettin, Wiltfang Jens, Otto Markus
Department of Neurology, University of Ulm, Germany.
Electrophoresis. 2015 Jan;36(2):355-62. doi: 10.1002/elps.201400339. Epub 2014 Dec 17.
The detection of oligoclonal bands (OCBs) in cerebrospinal fluid is an indicator of intrathecal synthesis of immunoglobulins which is a neurochemical sign of chronic inflammatory brain diseases. Intrathecally synthesized IgGs are typically observed in patients with multiple sclerosis. The current standard protocol for the detection of OCBs is IEF on agarose or polyacrylamide gels followed by immunoblotting or silver staining. These methods are time consuming, show substantial interlaboratory variation and cannot be used in a high throughput-approach. We have developed a new nanoscale method for the detection of OCBs based on automated capillary IEF followed by immunological detection. Evidence for intrathecal IgG synthesis was found in all tested patients (n = 27) with multiple sclerosis, even in two subjects who did not have oligoclonal bands according to standard methods. The test specificity was at 97.5% (n = 19). Our findings indicate that the novel OCB-CIEF-immunoassay is suitable for the rapid and highly sensitive detection of OCBs in clinical samples. Furthermore, the method allows for a higher sample throughput than the current standard methods.
脑脊液中寡克隆区带（OCBs）的检测是免疫球蛋白鞘内合成的一个指标，而免疫球蛋白鞘内合成是慢性炎症性脑疾病的一种神经化学标志。鞘内合成的IgG通常在多发性硬化症患者中观察到。目前检测OCBs的标准方案是在琼脂糖或聚丙烯酰胺凝胶上进行等电聚焦（IEF），然后进行免疫印迹或银染。这些方法耗时，实验室间差异很大，且不能用于高通量检测。我们开发了一种基于自动毛细管IEF随后进行免疫检测的检测OCBs的新纳米级方法。在所有测试的多发性硬化症患者（n = 27）中均发现了鞘内IgG合成的证据，甚至在两名根据标准方法没有寡克隆区带的受试者中也是如此。检测特异性为97.5%（n = 19）。我们的研究结果表明，新型OCB-CIEF免疫测定法适用于临床样本中OCBs的快速、高灵敏度检测。此外，该方法比当前标准方法具有更高的样本通量。
