Background: The international gold standard for avian influenza virus (AIV) diagnosis is virus isolation (VI) in specific pathogen-free embryonated chicken eggs (ECEs). AIV isolation typically involves a 6-day turnaround, during which premises under suspicion for notifiable AIV infection are held under restriction regardless of molecular diagnoses, often with significant welfare implications.

Methods: A reduction in time for negation by VI was investigated following experimental inoculation of AIV from known-positive original clinical material into ECEs. VI data derived from more than 600 case investigations from epizootics of high-pathogenicity AIV (HPAIV) in Great Britain since 2016 and from low-pathogenicity AIV (LPAIV) cases in Great Britain since 2014 were examined to support a reduction in test timing using alternative regimens.

Results: HPAIVs were isolated during the first passage, and for LPAIV VI, the second passage could be reduced to 2 days. Power analysis showed that the benefit of reducing the number of days outweighed the risk of missing a positive isolate.

Limitations: Limited data were available from experimental inoculations.

Conclusion: This truncated methodology, which enables an earlier release of restrictions, may substantially ease the economic implications of restriction. It could also reduce bird welfare implications and improve international standards without loss of test performance.