期刊
JOURNAL OF VIROLOGICAL METHODS
卷 127, 期 2, 页码 154-164出版社
ELSEVIER
DOI: 10.1016/j.jviromet.2005.04.007
关键词
BSE; immuno-PCR; IPCR; real-time IPCR; prion; PrPC; PrPSc; scrapie; TSE
Pathologic prion protein (PrPSc), implicated in transmissible spongiform encephalopathies, is detected by antibody-based tests or bioassays to confirm the diagnosis of prion diseases. Presently, the Western blot or an ELISA is officially used to screen the brain stem in cattle for the presence of PrPSc. The immuno-polymerase chain reaction (IPCR), a technique whereby the exponential amplification ability of PCR is coupled to the detection of proteins by antibodies in an ELISA format, was applied in-a modified real-time IPCR method to detect ultra-low levels of prion protein. Using IPCR, recombinant hamster PrPC was consistently detected at I fg/mL and proteinase K (PK)-digested scrapie infected hamster brain homogenates diluted to 10(-8) (approximately 10-100 infectious units) was detected with a semi-quantitative dose response. This level of detection is I million-fold more sensitive than the levels detected by Western blot or ELISA and poises IPCR as a method capable of detecting PrPSc in the pre-clinical phase of infection. Further, the data indicate that unless complete PK digestion of PrPC in biological materials is verified, ultrasensitive assays such as IPCR may inaccurately classify a sample as positive. (c) 2005 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据