TEST SYSTEM FOR DETECTING DNA OF CAUSATIVE AGENT OF MANNHEIMIOSIS (MANNHEIMIA HAEMOLYTICA) IN BIOLOGICAL MATERIAL OF ANIMALS AND FODDERS USING REAL-TIME POLYMERASE CHAIN REACTION Russian patent published in 2024 - IPC C12Q1/68 

FIELD: veterinary microbiology.

SUBSTANCE: described is a test system for detecting DNA of the causative agent of mannheimiosis (Mannheimia haemolytica) in biological material of animals and fodders using a real-time polymerase chain reaction. Test system includes a buffer for polymerase chain reaction, a mixture for its implementation, consisting of deoxynucleoside triphosphates, oligonucleotide primers and fluorescent probes specific for an animal genome region and for an internal control sample; mixture of enzymes from DNA polymerase with antibodies inhibiting enzyme activity, TAQ POLYMERASE, internal control sample in the form of a suspension of T4 bacteriophage with concentration of 5×10³ phage particles per 1 mcl, negative control sample, which is a mixture of recombinant plasmid DNA containing a fragment of the animal genome and a fragment of the genome of bacteriophage T4 with a nucleotide sequence: T4F: 5'-TACATATAAATCACGCAAAGC-3' — forward primer; TAR: 5'-TAGTATGGCTAATCTTATTGG-3' — reverse primer; T4P: CY5-5'-ACATTGGCACTGACCGAGTTC-3'-BHQ1 — probe, taken in volume ratio of 1:1. For the positive control sample, a fragment of the genome of the causative agent of mannheimiosis (Mannheimia haemolytica) with the following nucleotide sequence is used: Mh-F CTAACCAGGACAACCCACTA, Mh-R AGTTTAAGTAGYAAGCGTGTTCC, Mh-P ROX-CTAAGACTAAAATCGGATAGCCTGAAACGC-BHQ2; fluorescent dye ROX/Orange is used for DNA of the causative agent of mannheimiosis (Mannheimia haemolytica).

EFFECT: invention broadens functional capabilities and increases accuracy in detecting the causative agent of mannheimiosis (Mannheimia haemolytica).

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