Comparison of false-negative rates and limits of detection following macrofoam-swab sampling of Bacillus anthracis surrogates via Rapid Viability PCR and plate culture

Hutchison, J. R.; Piepel, G. F.; Amidan, B. G.; Hess, B. M.; Sydor, M. A.; Deatherage Kaiser, B. L.

doi:10.1111/jam.13706

Comparison of false-negative rates and limits of detection following macrofoam-swab sampling of Bacillus anthracis surrogates via Rapid Viability PCR and plate culture

Journal Article · Tue Mar 13 00:00:00 EDT 2018 · Journal of Applied Microbiology

DOI:https://doi.org/10.1111/jam.13706· OSTI ID:1434849

Hutchison, J. R. ^[1]; Piepel, G. F. ^[1]; Amidan, B. G. ^[1]; Hess, B. M. ^[1]; Sydor, M. A. ^[1]; Deatherage Kaiser, B. L. ^[1]

National Security Directorate, Pacific Northwest National Laboratory, Richland WA USA

Aims: We evaluated the effects of Bacillus anthracis surrogates, low surface concentrations, surface materials, and assay methods on false-negative rate (FNR) and limit of detection (LOD95) for recovering Bacillus spores using a macrofoam-swab sampling procedure. Methods and Results: Bacillus anthracis Sterne or Bacillus atrophaeus Nakamura spores were deposited over a range of low target concentrations (2 – 500 coupon-1) onto glass, stainless steel, vinyl tile, and plastic. Samples were assayed using a modified Rapid Viability-PCR (mRV-PCR) method and the traditional plate culture method to obtain FNR and LOD95 results. Conclusions: Mean FNRs tended to be lower for mRV-PCR compared to culturing, and increased as spore concentration decreased for all surface materials. Surface material, but not B. anthracis surrogate, influenced FNRs with the mRV-PCR method. The mRV-PCR LOD95 was lowest for glass and highest for vinyl tile. LOD95 values overall were lower for mRV-PCR than for the culture method. Significance and Impact of Study: This study adds to the limited data on FNR and LOD95 for mRV-PCR and culturing methods with low concentrations of B. anthracis sampled from various surface materials by the CDC macrofoam-swab method. These are key inputs for planning characterization and clearance studies for low contamination levels of B. anthracis.

Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (US)

Sponsoring Organization:: USDOE

DOE Contract Number:: AC05-76RL01830

OSTI ID:: 1434849

Report Number(s):: PNNL-SA-118902; 400904120

Journal Information:: Journal of Applied Microbiology, Journal Name: Journal of Applied Microbiology Journal Issue: 5 Vol. 124; ISSN 1364-5072

Country of Publication:: United States

Language:: English

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Comparison of false-negative rates and limits of detection following macrofoam-swab sampling of Bacillus anthracis surrogates via Rapid Viability PCR and plate culture

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