Immunological assay for spongiform encephalopathies
OwnerMichael O'Connorinventor
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Abstract
The invention relates to a method of detecting Transmissable Spongiform Encephalopathies (TSE) in animals, particulary in animal carcasses, using an anti-PrPsc antibody in an immunological assay. Also disclosed is a diagnostic kit for detecting TSE comprising the same antibody.
Claims8
A method for detecting the putative agent for TSE in animals comprising taking a body tissue sample from an animal, reacting the sample in a immunological assay with a labelled antibody which is capable of reacting with PrPSC and determining the amount of labelled antibody bound to the sample.
A method as claimed in any preceding claim wherein the antibodies are prion specific antibodies raised against one or more of the following sequences:
- MVKSIGSWILVLFVVAMWSDVGLCKKRPKPGGGINTGGSRYPGO-44
- GSPGGNRYPPOGGGGWGQPHGGGWGQPHGGGWGQPHGGGQGQP-87
- GGGGWGOGGSHSOWNKPSKPPKTNMKHVAGAAAGAVVGGLGGY-131
- MLGSAMSSPLIHFGNDYEDRYTRENMYRYPNOVYYRPVDRYSNQNN-177.
A method as claimed in any preceding claim wherein the immunological assay may be a competitive assay in which a solid support, suitably a microtitre plate, is pre-coated with a carrier protein-peptide conjugate, the animal tissue sample and an anti-peptide antibody are added to the solid support, allowed to react and the support washed a labelled anti-(anti-peptide antibody) antibody is added allowed to react, washed, a signal reagent added and the signal read.
The method as claimed in any preceding claim wherein the animals are selected from cattle, sheep and pigs and the tissue sample is taken from a carcass of such animals.
A method as claimed in any preceding claim wherein a sample of CNS tissue, suitably a cross-section of spinal cord, is used for testing.
A test kit for the detection of TSE in animals comprising an anti-peptide antibody raised against a synthetic peptide sequence having the general formula:
- ti X-(R1-Lys-His-R2)-Ala-Gly-Ala-Ala-Ala-R3-Gly-Ala-Val-Val-Gly-Gly-Leu-Gly-Gly-Tyr-Met-Leu-Gly-Ser-Ala-Met-Ser-(Arg-Pro-R4-R5)-Y
- wherein R1 is an amino acid residue selected from Met, Leu and Phe;
- R2 is either Met or Val;
- R3 is Ala or is absent;
- R4 and R5 are independently an amino acid residue selected from Leu, Ile and Met;
- one or more residues within brackets maybe present or absent with the provisio that if they are present they are attached to the rest of the peptide in sequence;
- and X and Y may each independently be absent or independently be one or more additional amino acid residues.
Dependencies are inferred from the claim text.