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- Title
Sulfated Dextrans Enhance In Vitro Amplification of Bovine Spongiform Encephalopathy PrP<sup>Sc</sup> and Enable Ultrasensitive Detection of Bovine PrP<sup>Sc</sup>.
- Authors
Murayama, Yuichi; Yoshioka, Miyako; Masujin, Kentaro; Okada, Hiroyuki; Iwamaru, Yoshifumi; Imamura, Morikazu; Matsuura, Yuichi; Fukuda, Shigeo; Onoe, Sadao; Yokoyama, Takashi; Mohri, Shirou
- Abstract
Background: Prions, infectious agents associated with prion diseases such as Creutzfeldt-Jakob disease in humans, bovine spongiform encephalopathy (BSE) in cattle, and scrapie in sheep and goats, are primarily comprised of PrPSc, a proteaseresistant misfolded isoform of the cellular prion protein PrPC. Protein misfolding cyclic amplification (PMCA) is a highly sensitive technique used to detect minute amounts of scrapie PrPSc. However, the current PMCA technique has been unsuccessful in achieving good amplification in cattle. The detailed distribution of PrPSc in BSE-affected cattle therefore remains unknown. Methodology/Principal Findings: We report here that PrPSc derived from BSE-affected cattle can be amplified ultraefficiently by PMCA in the presence of sulfated dextran compounds. This method is capable of amplifying very small amounts of PrPSc from the saliva, palatine tonsils, lymph nodes, ileocecal region, and muscular tissues of BSE-affected cattle. Individual differences in the distribution of PrPSc in spleen and cerebrospinal fluid samples were observed in terminal-stage animals. However, the presence of PrPSc in blood was not substantiated in the BSE-affected cattle examined. Conclusions/Significance: The distribution of PrPSc is not restricted to the nervous system and can spread to peripheral tissues in the terminal disease stage. The finding that PrPSc could be amplified in the saliva of an asymptomatic animal suggests a potential usefulness of this technique for BSE diagnosis. This highly sensitive method also has other practical applications, including safety evaluation or safety assurance of products and byproducts manufactured from bovine source materials.
- Subjects
PRIONS; CREUTZFELDT-Jakob disease; PRION diseases in animals; BOVINE spongiform encephalopathy; SCRAPIE; VIRUS diseases; SPLEEN; CEREBROSPINAL fluid; NERVOUS system; GOATS
- Publication
PLoS ONE, 2010, Vol 5, Issue 10, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0013152