TEST SYSTEM FOR DETECTING DNA OF CAUSATIVE AGENT OF MORAXELLOSIS KPC (MORAXELLA BOVIS) IN BIOLOGICAL MATERIAL OF ANIMALS AND FEED USING POLYMERASE CHAIN REACTION IN REAL TIME Russian patent published in 2024 - IPC C12Q1/68 

FIELD: veterinary microbiology.

SUBSTANCE: test system for detecting the DNA of the causative agent of bovine moraxellosis (Moraxella bovis) in animal biological material using real-time polymerase chain reaction is described. The test system includes a buffer for carrying out a polymerase chain reaction, a mixture for carrying it out, consisting of deoxynucleoside triphosphates, oligonucleotide primers and fluorescent probes specific for a region of the animal's genome and for an internal control sample; a mixture of enzymes from DNA polymerase with antibodies that inhibit enzyme activity, TAQ POLYMERASE, an internal control sample in the form of a suspension of bacteriophage T4 with a 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 T4 bacteriophage genome with the following nucleotide sequence: T4F: 5` - TACATATAAATCACGCAAAGC -3` - is a direct primer, T4R: 5` - TAGTATGGCTAATCTTATTGG-3` - is a reverse primer, T4P: CY5-5` - ACATTGGCACTGACCGAGTTC -3`-BHQ1 - is a probe, taken in a volumetric ratio of 1:1. Moreover, according to the invention, a fragment of the genome of the causative agent of pasteurellosis (Pasteurella multocida) with the following nucleotide sequence is used for a positive control sample: (5`-3`): Pm-F AACGTCCAATCAGTTGCGCC; Pm-R AGTGGGCTTGTCGGTAGTC; Pm-P R6G-CTCAACACACCAAACTCTGCCCAAC – BHQ1; T4-F TACATATAAATCACGCAAACG; T4-R TAGTATGGCTAATCTTATTGG; T4-P FAM-CATTGGCACTGACCGAGTTC-BHQ1 for the DNA of the causative agent of pasteurellosis (Pasteurella multocida), a fluorescent dye — ROX/Orange — was used.

EFFECT: expanding functionality and increasing accuracy in identifying the causative agent of pasteurellosis (Pasteurella multocida).

1 cl, 4 tbl, 1 ex