Pyrosequencing — a robust method for detecting antiviral resistance in seasonal and 2009 A (H1N1) influenza A viruses circulating with Department of Defense populations


Resistance to antiviral agents in influenza A viruses has steadily increased globally. The Centers for Disease Control (CDC) has reported incidences of 100% adamantane resistance in some influenza A subtypes and >98% of previous seasonal A/H1N1 is resistant to Oseltamivir. Single nucleotide polymorphisms (SNPs) with matrix (M2) and neuraminidase (NA) genes correlate well with in vitro and in vivo evidence of antiviral resistance. Pyrosequencing can be optimized to produce short, targeted genetic sequences that contain non-synonymous nucleotide changes that correlate with resistance to antiviral chemotherapeutics. The US Air Force School of Aerospace Medicine has employed Pyrosequencing to detect molecular markers of antiviral resistance within M2 and NA genes of influenza A viruses collected from Department of Defense (DoD) populations. A quick, highly sensitive, and cost-effective alternative to Sanger sequencing, Pyrosequencing allows rapid detection of common SNPs associated with adamantane and neuraminidase inhibitor (NAI) resistance respectively. Monitoring antiviral susceptibility within DoD populations complements influenza surveillance efforts and allows timely information exchange with the Armed Forces Health Surveillance Center, DoD sentinel sites, public health agencies, and collaborative partners.

This presentation provides:

An overview of the PyroMark Pyrosequencing based detection platform
An introduction to the DoD sentinel-based influenza surveillance program
A description of efforts to incorporate assays to detect M2-blocker and NAI susceptibilities in seasonal A/H3N2 viruses into the DoD program