Leclere Edwige, Moussa Aly, Chouaf-Lakhdar Latifa, Coleman Anthony W, Seigneurin Jean-Marie, Perron Hervé, Bencsik Anna
Laboratoire de Virologie Médicale, Centre Hospitalier Universitaire A. Michalon, BP17X 38043 Grenoble, France.
Biochem Biophys Res Commun. 2008 Aug 1;372(3):429-33. doi: 10.1016/j.bbrc.2008.05.040. Epub 2008 May 19.
The use of streptomycin in the PrP(sc) detection procedures represents a new and attractive way to detect more PrP(sc), the best marker for the transmissible spongiform encephalopathies (TSEs). Actually, the streptomycin PrP(sc) aggregating property reported recently was established as beneficial in PrP(sc) detection using immunohistochemistry in diagnostic as well as in experimental conditions. The present study reports in details how to use advantageously this original streptomycin property in PrP(res) biochemical extraction and detection. Using TSE diagnostic brain material, specificity and increased sensitivity using streptomycin-treated samples were substantiated. Then an early sequential brain and spleen sampling (from 7 to 49 days post-inoculation) from C57Bl/6 mice inoculated intra-cerebrally or intra-peritoneally with C506M3 scrapie strain was analysed using streptomycin versus ultracentrifugation PrP(res) extraction. Whatever the inoculation route, streptomycin allowed earlier PrP(res) detection in spleen (7 d.p.i.), then in brain suggesting a stronger affinity of the infectious agent for the lymphoid compartment.
在朊病毒蛋白(PrP)(sc)检测程序中使用链霉素是一种检测更多PrP(sc)的新的且有吸引力的方法,PrP(sc)是传染性海绵状脑病(TSEs)的最佳标志物。实际上,最近报道的链霉素使PrP(sc)聚集的特性在诊断和实验条件下利用免疫组织化学进行PrP(sc)检测时被证实是有益的。本研究详细报道了如何在PrP(res)的生化提取和检测中有利地利用这种链霉素的原始特性。使用TSE诊断脑材料,证实了使用链霉素处理的样品具有特异性且灵敏度提高。然后,对经脑内或腹腔内接种C506M3羊瘙痒病株的C57Bl/6小鼠在接种后7至49天进行早期连续脑和脾取样,使用链霉素与超速离心法提取PrP(res)进行分析。无论接种途径如何,链霉素都能使在脾脏中更早检测到PrP(res)(接种后7天),然后在脑中检测到,这表明感染因子对淋巴组织具有更强的亲和力。