Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrP(Sc) and enable ultrasensitive detection of bovine PrP(Sc).
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 PrP(Sc), a protease-resistant misfolded isoform of the cellular prion protein PrP(C). Pro...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2010-10-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2949392?pdf=render |
| _version_ | 1818311539939082240 |
|---|---|
| author | Yuichi Murayama Miyako Yoshioka Kentaro Masujin Hiroyuki Okada Yoshifumi Iwamaru Morikazu Imamura Yuichi Matsuura Shigeo Fukuda Sadao Onoe Takashi Yokoyama Shirou Mohri |
| author_facet | Yuichi Murayama Miyako Yoshioka Kentaro Masujin Hiroyuki Okada Yoshifumi Iwamaru Morikazu Imamura Yuichi Matsuura Shigeo Fukuda Sadao Onoe Takashi Yokoyama Shirou Mohri |
| author_sort | Yuichi Murayama |
| collection | DOAJ |
| description | 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 PrP(Sc), a protease-resistant misfolded isoform of the cellular prion protein PrP(C). Protein misfolding cyclic amplification (PMCA) is a highly sensitive technique used to detect minute amounts of scrapie PrP(Sc). However, the current PMCA technique has been unsuccessful in achieving good amplification in cattle. The detailed distribution of PrP(Sc) in BSE-affected cattle therefore remains unknown.We report here that PrP(Sc) derived from BSE-affected cattle can be amplified ultra-efficiently by PMCA in the presence of sulfated dextran compounds. This method is capable of amplifying very small amounts of PrP(Sc) from the saliva, palatine tonsils, lymph nodes, ileocecal region, and muscular tissues of BSE-affected cattle. Individual differences in the distribution of PrP(Sc) in spleen and cerebrospinal fluid samples were observed in terminal-stage animals. However, the presence of PrP(Sc) in blood was not substantiated in the BSE-affected cattle examined.The distribution of PrP(Sc) is not restricted to the nervous system and can spread to peripheral tissues in the terminal disease stage. The finding that PrP(Sc) 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. |
| first_indexed | 2024-12-13T08:03:34Z |
| format | Article |
| id | doaj.art-b8a835d01b9341b9bf9d5d227fd7cc18 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T08:03:34Z |
| publishDate | 2010-10-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-b8a835d01b9341b9bf9d5d227fd7cc182022-12-21T23:54:22ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-10-0151010.1371/journal.pone.0013152Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrP(Sc) and enable ultrasensitive detection of bovine PrP(Sc).Yuichi MurayamaMiyako YoshiokaKentaro MasujinHiroyuki OkadaYoshifumi IwamaruMorikazu ImamuraYuichi MatsuuraShigeo FukudaSadao OnoeTakashi YokoyamaShirou MohriPrions, 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 PrP(Sc), a protease-resistant misfolded isoform of the cellular prion protein PrP(C). Protein misfolding cyclic amplification (PMCA) is a highly sensitive technique used to detect minute amounts of scrapie PrP(Sc). However, the current PMCA technique has been unsuccessful in achieving good amplification in cattle. The detailed distribution of PrP(Sc) in BSE-affected cattle therefore remains unknown.We report here that PrP(Sc) derived from BSE-affected cattle can be amplified ultra-efficiently by PMCA in the presence of sulfated dextran compounds. This method is capable of amplifying very small amounts of PrP(Sc) from the saliva, palatine tonsils, lymph nodes, ileocecal region, and muscular tissues of BSE-affected cattle. Individual differences in the distribution of PrP(Sc) in spleen and cerebrospinal fluid samples were observed in terminal-stage animals. However, the presence of PrP(Sc) in blood was not substantiated in the BSE-affected cattle examined.The distribution of PrP(Sc) is not restricted to the nervous system and can spread to peripheral tissues in the terminal disease stage. The finding that PrP(Sc) 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.http://europepmc.org/articles/PMC2949392?pdf=render |
| spellingShingle | Yuichi Murayama Miyako Yoshioka Kentaro Masujin Hiroyuki Okada Yoshifumi Iwamaru Morikazu Imamura Yuichi Matsuura Shigeo Fukuda Sadao Onoe Takashi Yokoyama Shirou Mohri Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrP(Sc) and enable ultrasensitive detection of bovine PrP(Sc). PLoS ONE |
| title | Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrP(Sc) and enable ultrasensitive detection of bovine PrP(Sc). |
| title_full | Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrP(Sc) and enable ultrasensitive detection of bovine PrP(Sc). |
| title_fullStr | Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrP(Sc) and enable ultrasensitive detection of bovine PrP(Sc). |
| title_full_unstemmed | Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrP(Sc) and enable ultrasensitive detection of bovine PrP(Sc). |
| title_short | Sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy PrP(Sc) and enable ultrasensitive detection of bovine PrP(Sc). |
| title_sort | sulfated dextrans enhance in vitro amplification of bovine spongiform encephalopathy prp sc and enable ultrasensitive detection of bovine prp sc |
| url | http://europepmc.org/articles/PMC2949392?pdf=render |
| work_keys_str_mv | AT yuichimurayama sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT miyakoyoshioka sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT kentaromasujin sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT hiroyukiokada sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT yoshifumiiwamaru sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT morikazuimamura sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT yuichimatsuura sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT shigeofukuda sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT sadaoonoe sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT takashiyokoyama sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc AT shiroumohri sulfateddextransenhanceinvitroamplificationofbovinespongiformencephalopathyprpscandenableultrasensitivedetectionofbovineprpsc |