Development of Avian and Human Influenza Analytical Reference Materials for Diagnostics and Surveillance

Human and highly pathogenic avian influenza (HPAI) viruses pose a significant public health risk due to their potential for widespread illness and economic consequences. Effective surveillance and diagnostic testing are crucial for early detection and control of outbreaks. ATCC® has developed a comprehensive suite of quantitative synthetic analytical reference materials (ARMs) for HPAI virus serotypes H5N1, H5N6, H7N7, H7N9, and H9N2; human influenza A virus serotypes H1N1, H3N2, and H1N1 2009 pandemic; and influenza B virus strains. Each synthetic ARM contains nearly complete gene sequences from segments 4, 5, 6, 7, and 8, including the HA, NP, NA, M1, M2, and NEP/NS1 genes, covering 50% of the influenza genome. These segments are key diagnostic targets for molecular tests and provide sufficient genomic context for assessing assay specificity. Manufactured using reliable synthetic biology technology, verified through next-generation sequencing, and quantitated via digital PCR, these ARMs do not contain any viable material and can be handled in BSL-1 settings. The synthetic ARMs were evaluated using several published quantitative PCR assays, including those from the Centers for Disease Control and Prevention, the World Health Organization, and the World Organization for Animal Health. The synthetic products displayed equal performance to genomic RNA during all tests. Our findings indicate that the synthetic ARMs are equivalent to their corresponding genomic RNA, making them a valuable BSL-1 alternative to BSL-3–derived materials. These synthetic ARMs can serve as reliable and safe controls for molecular assays used in diagnostics and surveillance.