Inactivation of Viable Surrogates for the Select Agents Virulent Newcastle Disease Virus and Highly Pathogenic Avian Influenza Virus Using Either Commercial Lysis Buffer or Heat.

INTRODUCTION

Federal Select Agent Program regulations require laboratories to document a validated procedure for inactivating select agents prior to movement outside registered space. Avian influenza viruses and virulent Newcastle disease virus (vNDV) are cultured in chicken amnio-allantoic fluid (AAF), but the efficacy of commercial lysis buffers to inactivate viruses in protein-rich media has not been documented.

OBJECTIVES

We assesses the efficacy of MagMAX™ lysis buffer for inactivating highly pathogenic avian influenza virus (HPAIV) and vNDV in chicken AAF and confirm the inactivation of avian influenza in serum using heat.

METHODS

Low pathogenic avian influenza virus (LPAIV) and avian paramyxovirus subtype-1 (APMV-1) were incubated with lysis buffer and tested for viability. Known viable LPAIV and APMV-1 RNA was extracted from AAF using MagMAX™-96 AI/ND Viral RNA Isolation kit, and the eluate was tested for remaining infectious agent. Finally, inactivation of LPAIV in serum was examined over 3 combinations of temperature and incubation time.

RESULTS

MagMAX™ lysis buffer inactivated both LPAIV and APMV-1 in AAF when incubated for 30 minutes at room temperature. The full extraction process eliminated viable virus from the final RNA eluate. LPAIV in serum heated to 70°C for 30 minutes was rendered noninfectious.

CONCLUSION

The ability of a diagnostic laboratory to move samples from one space to another is critical to maintaining biosecurity as well as efficient laboratory workflow. Our study demonstrates a method to ensure the inactivation of viable avian influenza and avian paramyxoviruses in AAF, RNA eluate, and viable avian influenza virus in sera.