TEST SYSTEM OF DETECTING DNA OF AFRICAN SWINE FEVER VIRUS BY REAL-TIME POLYMERASE CHAIN REACTION Russian patent published in 2018 - IPC C12Q1/68 

FIELD: veterinary medicine.

SUBSTANCE: invention relates to the field of veterinary virology, in particular to a test system for detecting the DNA of a particularly dangerous agent of African swine fever (ASF). Test system for detecting DNA of the African swine fever virus using real-time polymerase chain reaction includes plastic vials and test tubes, a thermostable Tag-polymerase enzyme, a buffer for the reaction, a mixture of four deoxynucleotide triphosphates, a positive control – recombinant plasmid containing a fragment of the vp72 gene of the African swine fever virus, synthetic oligonucleotide primers and a probe. Synthetic oligonucleotide primers and a fluorescent probe complementary to the preserved region of the genome of the African swine fever virus gene vp72 region are used and have the following nucleotide composition: SEQ NO 1: 5’-CTG-CTC-ATG-GTA-TCA-ATC-3’ – forward primer, SEQ NO 2: 5’-GAT-ACC-ACA-AGA-TCG-CCG-3’ – reverse primer, SEQ NO 3: 5’-FAM-CCA-CGG-GAG-GAA-TAC-CAA-CCC-AGT-G-BHQ1-3’ – a fluorescent probe and taken at a ratio of 1:1:0.5, wherein BHQ1 – a dark fluorescence quenching agent is attached to 3'-terminal nucleotide, and FAM – a fluorescent dye is attached to nucleotide C, where a 5-fold buffer is used to set up the reaction and distilled water is used as the negative control.

EFFECT: invention is intended to increase the degree of specificity and sensitivity of the test system, as well as to shorten the duration of diagnosis while reducing the likelihood of technological errors during laboratory manipulations.

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