Quadrio Isabelle, Ugnon-Café Séverine, Dupin Maryline, Esposito Guillaume, Streichenberger Nathalie, Krolak-Salmon Pierre, Vital Anne, Pellissier Jean-François, Perret-Liaudet Armand, Perron Hervé
Neurobiology Department, Groupement Hospitalier Est, Hospices Civils de Lyon, Lyon, France.
Lab Invest. 2009 Apr;89(4):406-13. doi: 10.1038/labinvest.2008.169. Epub 2009 Feb 2.
The use of streptomycin in the pathological prion protein (PrP(sc)) detection procedures represents a new and attractive way for diagnostic purpose. With this agent, western blot readily detected PrP(sc) in 263K scrapie hamster and C57Bl/6 wild-type mice challenged with C506M3 scrapie strain. Our aim was to evaluate this new diagnosis procedure in the field of human transmissible spongiform encephalopathies (TSEs). First, we had confirmed the ability of streptomycin to precipitate PrP(res) from human brain of Creutzfeldt-Jakob disease (CJD) patient. Second, we compared the detection of PrP(res) with streptomycin against three other protocols using other precipitations. Then we assessed PrP(res) detection with streptomycin in 98 brain tissue samples from various aetiologies of human TSEs and 52 brain samples from other dementia. Finally, we applied this protocol for tonsils examination of five patients suspected of variant CJD (v-CJD). Sensitivity and specificity obtained with the streptomycin protocol were both 100% on brain tissue. For tonsil tissues, PrP(res) was clearly identified in the two post-mortem confirmed v-CJD cases, whereas no characteristic three-band pattern was seen in the three confirmed non-v-CJD samples. In this study, streptomycin demonstrated its efficiency to detect PrP(res) both in the central nervous system and in the lymphoid tissue without practical difficulty and with rapid preparation. Because of its ability to act as a good agent for PrP(sc) examination in different tissues, recovery of PrP(sc) in biological fluids using streptomycin should open further perspectives of applications in CJD diagnostics. Streptomycin effects in vivo might thus also be questioned.
在病理性朊病毒蛋白（PrP(sc)）检测程序中使用链霉素代表了一种用于诊断目的的新的且有吸引力的方法。使用这种试剂，蛋白质免疫印迹法能轻易检测出用C506M3羊瘙痒病株攻击的263K羊瘙痒病仓鼠和C57Bl/6野生型小鼠中的PrP(sc)。我们的目的是在人类传染性海绵状脑病（TSEs）领域评估这种新的诊断程序。首先，我们已证实链霉素能够从克雅氏病（CJD）患者的人脑中沉淀出PrP(res)。其次，我们将用链霉素检测PrP(res)与使用其他沉淀方法的另外三种方案进行了比较。然后我们评估了在98份来自各种人类TSE病因的脑组织样本和52份来自其他痴呆症的脑样本中用链霉素检测PrP(res)的情况。最后，我们将该方案应用于对五名疑似变异型CJD（v-CJD）患者的扁桃体检查。在脑组织上，链霉素方案获得的敏感性和特异性均为100%。对于扁桃体组织，在两例死后确诊的v-CJD病例中清楚地鉴定出了PrP(res)，而在三例确诊的非v-CJD样本中未观察到特征性的三条带模式。在本研究中，链霉素证明了其在中枢神经系统和淋巴组织中检测PrP(res)的效率，且没有实际困难且制备迅速。由于其能够作为不同组织中PrP(sc)检测的良好试剂，使用链霉素在生物体液中回收PrP(sc)应该为CJD诊断开辟进一步的应用前景。因此，链霉素在体内的作用也可能受到质疑。
