| 摘要 |
<p>#CMT# #/CMT# In vitro method of detecting a pathogenic conformer of a prion protein (PrPSc) in a sample, comprises: (a) contacting coated nanoshells of plasminogen with a sample; (b) separating the coated nanoshells of plasminogen carrying PrPSc from the sample; (c1) contacting the coated nanoshells of plasminogen carrying PrPSC obtained in step (b) with a preparation comprising non-pathogenic conformer of the prion protein (PrPC); (c2) disintegrating the aggregates optionally formed during step (c1); and (d) detecting the presence of PrPsc in the preparation. #CMT# : #/CMT# In vitro method of detecting a pathogenic conformer of a prion protein (PrPSc) in a sample, comprises: (a) contacting coated nanoshells of plasminogen with a sample; (b) separating the coated nanoshells of plasminogen carrying PrPSc from the sample; (c1) contacting the coated nanoshells of plasminogen carrying PrPSC obtained in step (b) with a preparation comprising non-pathogenic conformer of the prion protein (PrPC); (c2) disintegrating the aggregates optionally formed during step (c1); and (d) detecting the presence of PrPsc in the preparation, where the presence of PrPsc in the preparation is indicative of the presence of PrPsc in the sample and the steps (c1) and (c2) form a cycle which is repeated at least twice before implementing step (d). Independent claims are included for: (1) kit comprising paramagnetic nanoshells, plasminogen, optionally at least one other ligand of a pathogenic conformer of the prion protein and a known amount of non-pathogenic conformer of the prion protein and optionally instructions for implementing the method; and (2) a method for decontaminating a biological sample to contain pathogenic conformer of the prion protein (PrPSc), comprising contacting the coated paramagnetic nanoshells of plasminogen with a sample, separating the coated nanoshells of plasminogen sample and collecting the decontaminated sample. #CMT#USE : #/CMT# The method is useful for detecting a pathogenic conformer of a prion protein in a sample, decontaminating a biological sample liable to contain pathogenic conformer of the prion protein (claimed) and diagnosing the diseases in patients. #CMT#ADVANTAGE : #/CMT# The method detects pathogenic conformers of prion diseases with high sensitivity thus accurately diagnosing the diseases. #CMT#BIOLOGY : #/CMT# Preferred Components: The nanoshells are coated paramagnetic nanoshells of plasminogen in an amount of 10-30 mu g of plasminogen/mg of nanoshells. #CMT#INSTRUMENTATION AND TESTING : #/CMT# Preferred Method: The step (d) comprises (d1) destruction or removal of the PrPC protein present in the preparation after step (c2). The coated nanoshells of plasminogen are contacted with the sample in step (a) at a ratio of 2.5-90 mu l of nanoshells suspension (1 wt./vol.%) for 50-500 mu l of sample. The step (a) is performed with stirring and the disintegration of step (c2) is carried out by sonication. The method comprises (a) contacting the coated paramagnetic plasminogen nanoshells with a sample, in an amount of from 10-30 mu g plasminogen/mg of nanoshells. The step (c1) is carried out for 20 minutes to 2 hours. The step (c2) is carried out for 10 seconds to 1 minute. The step (c1) is carried out for 60-120 minutes, preferably 80-100 minutes during a first amplification cycle. The step (c2) is carried out by sonication for 10 seconds to 1 minutes, preferably 20-40 seconds. The step (c1) is carried out for 30-60 minutes during one or more amplification cycles, preferably 49-99 amplification cycles. The steps (c1) and (c2) form a cycle which is carried out 50-350 times before implementing step (d). The method comprises: detecting the presence of PrPsc in step (d) which is indicative of a transmissible spongiform encephalopathy disease in the subject, where the sample is a biological sample from a subject; detecting the presence of PrPsc in step (d) which is indicative of a liquid or solid food contaminated with pathogenic conformer of PrPSC, where the sample is a liquid or solid food; and detecting the presence of PrPsc in step (d) which is indicative of inappropriate blood or plasma transfusion or inappropriate organ transplantation, where the sample is, or is derived from, a sample of blood, plasma or organ and the transmissible spongiform encephalopathy is Creutzfeldt-Jakob disease or its variant. The step (d) comprises quantizing the pathogenic conformer.</p> |
