IMMUNO-PCR DIAGNOSTIC TECHNIQUE FOR INFECTIOUS SPONGIFORM ENCEPHALOPATHY IN ANIMALS Russian patent published in 2013 - IPC G01N33/48 C12N15/00 

FIELD: medicine.

SUBSTANCE: technique involves the stages of immobilising an antigen into the wells of a polystyrene 96-well plate, binding the antigen to a biotylated antigen, conducting a reaction of the biotylated antigen and a streptavidin complex (a complex of streptavidin - biotin - DNA target), conducting a real-time polymerase chain reaction using a fluorescent probe. What is used is the DNA target with a sequence: ACCAOCTCCCCACCACTOCCAACCACCTCOCOTA GGATAGAGTCAGTCCTTGGCCTCCTTGGCCCAGTTAAGAAGTTGCAGCC ACACACGCTGTTGTTGGGTTCGGGGCGGAGTTGCAGCCATCTACACAA ACGATACCCTCGTGCAGCTGGAGAAGCAGCACGGCCTATTACCTGGAG GAGGATCGAAACTGA, and the monoclonal antibody 15B3. The antibody reacts with a pathogenic prion protein, but not with a normal one, and detects PrP^Sc in homogenates of the infected material without the need of action of proteinases. The technique enables simplifying the real-time immuno-PCR method and increasing its sensitivity by immobilising the antigen on a plate substrate with using no capture antibodies, as well as using the DNA target having no homology with the GenBank database sequences.

EFFECT: reducing a risk of false responses as a result of exogenous contamination.

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