METHOD FOR DETERMINING CONCENTRATION OF VIRIONS OF CAUSATIVE AGENT OF CANINE PARVOVIRAL ENTERITIS IN RAW MATERIAL FOR VACCINES Russian patent published in 2024 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: described is a method for indirect determination of the concentration of virions of the causative agent of canine parvoviral enteritis in raw material for vaccines. Method involves: recovering DNA of the causative agent of canine parvoviral enteritis by a solid-phase method; conducting an amplification reaction in real time with detection of a fluorescent sigmoid signal for the analysed samples using oligonucleotides designed for the target region of the NS₁ gene of the causative agent of canine parvoviral enteritis in range of 381…638 bp; calculation of the maximum on the graph of the second derivative of the sigmoid function for the products of the amplification reaction; determination of the infectious activity titre of the causative agent of canine parvoviral enteritis in the raw material for the vaccine based on the developed mathematical function C_CPV = -0.3012 × C_p-NS1 + 9.0213 with high accuracy of approximation R² = 0.9944 and amplification efficiency of 99.74%. In the invention, dependence of the concentration of CPV virions and values of points C_p-NS1 are established with high accuracy of approximation (R² = 0.9944) and amplification efficiency of 99.74%, which enables to indirectly determine the concentration of virions of the causative agent of canine parvoviral enteritis in the raw material for vaccines using the amplification of the NS₁-gene of viral DNA.

EFFECT: main advantages of the invention are the possibility of eliminating the probability of contamination during the reaction and subjectivity when taking into account the analysis results; possibility to reduce time of analysis of virus-containing raw material for vaccines to 2 hours; possibility to increase specificity and sensitivity of analysis.

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