T/CVDA 1-2022 Active

Scope: This standard specifies the experimental operation methods and result determination criteria for the plaque reduction neutralization test used to measure neutralizing antibodies in animal serum against West Nile virus. This standard is applicable to the qualitative and quantitative detection of neutralizing antibodies against West Nile virus in animal serum. Main technical content: This technical standard is suitable for the detection of neutralizing antibodies against West Nile virus in serum of animals susceptible to the virus. By detecting neutralizing antibodies against West Nile virus in serum, it can assist in determining whether an animal is infected with West Nile virus or evaluating the immunization effect of the West Nile fever vaccine. The main technical content includes: 1. Construction of a cell line stably expressing the West Nile virus C-prM-E protein; 2. Construction of a West Nile virus replicon lacking the cellular protein gene and expressing green fluorescent protein; 3. Packaging and titration of the West Nile virus replicon; 4. Collection and inactivation of animal serum; 5. Neutralization of serum with the West Nile virus replicon and cell inoculation; 6. Observation and determination of results through fluorescent foci; 7. Observation and determination of results through virus plaque count.