Applied Neurobiology Group, Division of Veterinary Bioscience, School of Veterinary Medicine, University of Glasgow, Bearsden Road, Glasgow G61 1QH, UK; Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, University Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Vet J. 2013 Sep;197(3):836-41. doi: 10.1016/j.tvjl.2013.05.039. Epub 2013 Jun 29.
Cerebrospinal fluid (CSF) is a potential source for disease-specific biomarkers that may assist in the staging and determining the prognosis of neurodegenerative conditions in animals. However, the validity of such putative biomarkers may be influenced by pre-analytical variables, including the procedures adopted to collect and store the CSF. This study assessed the effect of three handling practices on the stability of a panel of CSF proteins: clusterin (also known as apolipoprotein J), haptoglobin, cystatin C, and transthyretin (TTR). The three handling procedures for canine CSF were mimicked in the laboratory as follows: (1) storage in a refrigerator overnight (4 °C for 18 h); (2) carrying a sample in the pocket of a clinician (37 °C for 4h); and (3) mailing a sample to a remote laboratory for analysis (room temp for 48 h). The impact of these three scenarios on the concentrations of the selected proteins was assessed using Western blotting and compared to an aliquot of CSF that had been kept frozen. The level of clusterin was significantly reduced following 48 h at room temperature (P<0.05), while the concentration of the dimeric form of TTR increased following this handling procedure and also when held at 37 °C for 4h. A reducing agent prevented this increase at 37 °C. In conclusion, exposing CSF samples to various environmental conditions can significantly alter their protein content, a factor that must be considered in studies assessing potential biomarkers in canine CSF.
脑脊液（CSF）是疾病特异性生物标志物的潜在来源，可能有助于对动物的神经退行性疾病进行分期和预后判断。然而，这种假定的生物标志物的有效性可能受到分析前变量的影响，包括用于采集和储存 CSF 的程序。本研究评估了三种处理方法对一组 CSF 蛋白稳定性的影响： 簇蛋白（也称为载脂蛋白 J）、触珠蛋白、胱抑素 C 和转甲状腺素（TTR）。在实验室中模拟了犬 CSF 的三种处理程序，如下所示：（1）在冰箱中过夜储存（4°C 18 h）；（2）在医生口袋中携带样本（37°C 4h）；（3）将样本邮寄到远程实验室进行分析（室温 48 h）。使用 Western blot 评估这三种情况对选定蛋白质浓度的影响，并与冷冻保存的 CSF 等分试样进行比较。在室温下放置 48 小时后，簇蛋白的水平显着降低（P<0.05），而 TTR 的二聚体形式的浓度在经过这种处理程序后以及在 37°C 下放置 4 小时后增加。在 37°C 时使用还原剂可防止这种增加。总之，将 CSF 样本暴露于各种环境条件下会显著改变其蛋白质含量，这是在评估犬 CSF 中潜在生物标志物的研究中必须考虑的因素。
