Comparative analysis of the extraction methods of avian influenza virus from raw cow's mil

The article presents data on the epizootic situation of avian influenza (AI) in Russia and in the world for the general assessment of the risks of outbreaks of AI in cattle in Russia, as well as the results of testing the effectiveness of the methods of virus RNA isolation from milk for molecular di agnostics. There has been a trend of a global increase in the number of AI outbreaks not only in birds but also in mammals, including humans. Of particular concern to the scientific community is the out break of highly pathogenic avian influenza virus (HPAI) subtype H5N1 in cattle in the United States reported in February 2024. Since notification began, the total number of infected animals has exceeded 80 animals. In this situation, the potential threat to human health is posed by the presence of the virus in high concentrations in cow's milk. The study was conducted in 2024. Data on the AI outbreaks from the World Animal Health Information System were used to describe the epizootic pattern of the HPAI virus. Laboratory studies have been conducted using commercial kits designed for RNA extraction from biological material based on nucleic acid sorption on silica gel surface and magnetic separation available on the Russian market. The efficiency of RNA isolation was evaluated by real-time reverse transcription polymerase chain reaction (RT-PCR) method. The data of the epizootic situation on AI indicate the need for careful monitoring of the virus spread among birds and mammals. Despite the low risk of Influenza A detection among cattle in Russia, it is necessary to maintain a high level of preparedness for diagnostic tests. A commercial kit for RNA isolation based on magnetic sorption followed by reverse transcription with a commercial kit “REVERTA-L” and RT-PCR according to FLI protocol, Germany (Friedrich Loeffler Institute) showed optimal results for milk testing for AI.

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