Rapid and Highly Sensitive Detection of Variant Creutzfeldt - Jakob Disease Abnormal Prion Protein on Steel Surfaces by Protein Misfolding Cyclic Amplification: Application to Prion Decontamination Studies
Rapid and Highly Sensitive Detection of Variant Creutzfeldt - Jakob Disease Abnormal Prion Protein on Steel Surfaces by Protein Misfolding Cyclic Amplification: Application to Prion Decontamination Studies
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DOI：
10.1371/journal.pone.0146833
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发表时间：
2016-01-22
期刊：
影响因子：
3.7
通讯作者：
Bougard, Daisy
中科院分区：
文献类型：
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作者：
Belondrade, Maxime;Nicot, Simon;Bougard, Daisy
尽管最近一项对存档阑尾组织的调查估计，英国每 2000 人中就有 1 人异常朊病毒蛋白 (PrPTSE) 呈阳性，但人群中变异型克雅氏病 (vCJD) 的患病率仍不确定。 vCJD 朊病毒显着的向淋巴细胞性增加了医疗机构中某些外科手术可能存在医源性 vCJD 传播风险的可能性。因此，在医疗器械再处理之前对其应用的净化程序进行彻底验证至关重要。目前的限制是缺乏允许人类朊病毒的快速模型。在这里，我们开发了一种基于蛋白质错误折叠循环扩增（PMCA）技术并结合不锈钢丝表面作为朊病毒载体的朊病毒检测方法（Surf-PMCA）。该测定可以在两周的时间内特异性检测到单根钢丝上吸附的微量（10(-8) 脑稀释液）人类 vCJD 或绵羊痒病 PrPTSE。使用 Surf-PMCA，我们评估了几种参考和市售的朊病毒特异性净化程序的性能。令人惊讶的是，我们发现几种市售试剂去除人 vCJD PrPTSE 的效率低于预期。总体而言，我们的结果表明，Surf-PMCA 可用作快速、超灵敏的检测方法，用于检测吸附在金属表面上的人 vCJD PrPTSE，从而促进针对人朊病毒的净化程序的开发和验证。
The prevalence of variant Creutzfeldt-Jakob disease (vCJD) in the population remains uncertain, although it has been estimated that 1 in 2000 people in the United Kingdom are positive for abnormal prion protein (PrPTSE) by a recent survey of archived appendix tissues. The prominent lymphotropism of vCJD prions raises the possibility that some surgical procedures may be at risk of iatrogenic vCJD transmission in healthcare facilities. It is therefore vital that decontamination procedures applied to medical devices before their reprocessing are thoroughly validated. A current limitation is the lack of a rapid model permissive to human prions. Here, we developed a prion detection assay based on protein misfolding cyclic amplification (PMCA) technology combined with stainless-steel wire surfaces as carriers of prions (Surf-PMCA). This assay allowed the specific detection of minute quantities (10(-8) brain dilution) of either human vCJD or ovine scrapie PrPTSE adsorbed onto a single steel wire, within a two week timeframe. Using Surf-PMCA we evaluated the performance of several reference and commercially available prion-specific decontamination procedures. Surprisingly, we found the efficiency of several marketed reagents to remove human vCJD PrPTSE was lower than expected. Overall, our results demonstrate that Surf-PMCA can be used as a rapid and ultrasensitive assay for the detection of human vCJD PrPTSE adsorbed onto a metallic surface, therefore facilitating the development and validation of decontamination procedures against human prions.