Highly infectious purified preparations of disease-specific amyloid of transmissible spongiform encephalopathies are not devoid of nucleic acids of viral size.
An efficient purification protocol for infectivity causing a transmissible spongiform encephalopathy (TSE) is described. From fractions purified by this protocol about 3 x 10(8) LD50 but only 3 ng of nucleic acids per gram of brain material can be isolated from all TSE-affected brains (hamster, human, sheep, cattle). By PAGE such fractions from brains of infected and control hamsters contained only one distinct nucleic acid band of 1.5 kg together with some broader smear of nucleic acid material. Although distilled water was used for such purifications, quite often a similar nucleic acid band was isolated from blanks containing no brain material. In all instances this material proved to be DNA. The result challenges the potentially important claim that purified infectious preparations of TSE-specific amyloid are free of nucleic acids of viral size. Nucleic acids isolated by other groups from diseased brain were not detected in preparations isolated by the new protocol. The application of this purification protocol in future studies will be helpful to decide whether TSEs are caused by agents containing nucleic acid or by protein only.
Diringer
H
H
Department of Virology, Robert-Koch-Institut, Berlin, Germany.
Beekes
M
M
Ozel
M
M
Simon
D
D
Queck
I
I
Cardone
F
F
Pocchiari
M
M
Ironside
J W
JW
D000682:Amyloid / Q000302:isolation & purification*; D000818:Animals; D002417:Cattle; D006224:Cricetinae; D004247:DNA / Q000302:isolation & purification*; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005069:Evaluation Studies as Topic; D006801:Humans; D010316:Particle Size; D017096:Prion Diseases / Q000209:etiology*; D011506:Proteins / Q000302:isolation & purification; D012313:RNA / Q000302:isolation & purification; D012756:Sheep
D000682:Amyloid; D004279:DNA, Viral; D011506:Proteins; D012313:RNA; D004247:DNA