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. 2019 Dec 1;24(4):189-199.
doi: 10.1177/1535676019888920. Epub 2019 Dec 1.

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

Katrina Alger  1 , Hon Ip  1 , Jeffrey Hall  1 , Sean Nashold  1 , Katherine Richgels  1 , Carrie Smith  1
Affiliations

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

Katrina Alger et al. Appl Biosaf. .

Abstract

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.

Keywords: BSL-3; Federal Select Agent Program; biological select agents and toxins; biorisk management; infectious agent; pathogen.

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Figures

Figure 1.
Figure 1.
Illustrated workflow for MagMAX™ lysis buffer inactivation of APMV-1 and H5N2 LPAIV in AAF. (a) Workflow for initial trial (APMV-1 only) using variable incubation times at room temperature, prior to dilution with VTM and inoculation into 8-day old embryonated chicken eggs. (b) Workflow for all subsequent trials (APMV-1 and H5N2 LPAIV) adding 1 minute of agitation on an orbital shaker, followed by 30 minutes of incubation at room temperature, prior to dilution with VTM and inoculation into 8-day old embryonated chicken eggs. AAF, amnio-allantoic fluid; APMV-1, avian paramyxovirus subtype-1; LPAIV, low pathogenic avian influenza; VTM, viral transport media.
Figure 2.
Figure 2.
Illustrated workflow for egg inoculation in MagMAX™ lysis buffer inactivation trials (avian paramyxovirus subtype-1 and H5N2 low pathogenic avian influenza).
See this image and copyright information in PMC

References

    1. Centers for Disease Control and United States Department of Agriculture. Federal Select Agent Program. 2017. https://www.selectagents.gov. Accessed September 16, 2017.
    1. Centers for Disease Control and United States Department of Agriculture. Agriculture: possession, use, and transfer of select agents and toxins. Fed Regist. 2005;70 FR 13278. Codified at 7 CFR § 331.
    1. Centers for Disease Control and United States Department of Agriculture. Animal and animal products: possession, use, and transfer of select agents and toxins. Fed Regist. 2005;70 FR 13284. Codified at 9 CFR § 121.
    1. Centers for Disease Control and United States Department of Agriculture. Public health: possession, use, and transfer of select agents and toxins. Fed Regist. 2005;70 FR 13316. Codified at 42 CFR § 73.
    1. United States Department of Health and Human Services, Public Health Service. Biosafety in microbiological and biomedical laboratories. 5th ed. 2009. HHS Pub No. (CDC 21-1112). https://www.cdc.gov/biosafety/publications/bmbl5/BMBL.pdf. Accessed September 17, 2017.

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