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. Early detection and control of outbreaks rely on effective surveillance and diagnostic testing. 

ATCC® 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 the complete sequences from segments 4, 5, 6, 7, and 8, including the HA, NP, NA, M1, M2, NS1, 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. These ARMs are manufactured using a highly reliable synthetic biology technology, verified through next-generation sequencing, and quantified via Droplet Digital PCR (Bio-Rad Laboratories, Inc.). Furthermore, they do not contain any viable material and can be handled in a BSL-1 setting. As such, they are intended to serve as safe and reliable positive controls for molecular tests for surveillance and diagnostics. 

The synthetic ARMs were experimentally evaluated using several published quantitative PCR assays, including those from the Centers for Disease Control and Prevention, the World Health Organization, the World Organization for Animal Health, and other highly cited sources. We conducted an in silico assessment of ARM compatibility with over 250 publicly available published assays. The synthetic products displayed equal performance to genomic RNA during all tests.