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Title: Evaluation of PCR Systems for Field Screening of Bacillus anthracis

Abstract

There is little published data on the performance of hand-portable polymerase chain reaction (PCR) instruments that could be used by first responders to determine if a suspicious powder contains a potential biothreat agent. We evaluated five commercially available hand-portable PCR instruments for detection of Bacillus anthracis (Ba). We designed a cost-effective, statistically-based test plan that allows instruments to be evaluated at performance levels ranging from 0.85-0.95 lower confidence bound (LCB) on the probability of detection (POD) at confidence levels of 80-95%. We assessed specificity using purified genomic DNA from 13 Ba strains and 18 Bacillus near neighbors, interference with 22 common hoax powders encountered in the field, and PCR inhibition when Ba spores were spiked into these powders. Our results indicated that three of the five instruments achieved >0.95 LCB on the POD with 95% confidence at test concentrations of 2,000 genome equivalents/mL (comparable to 2,000 spores/mL), displaying more than sufficient sensitivity for screening suspicious powders. These instruments exhibited no false positive results or PCR inhibition with common hoax powders, and reliably detected Ba spores spiked into common hoax powders, though some issues with instrument controls were observed. Our testing approach enables efficient instrument performance testing to a statistically rigorousmore » and cost-effective test plan to generate performance data that will allow users to make informed decisions regarding the purchase and use of biodetection equipment in the field.« less

Authors:
; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1344637
Report Number(s):
PNNL-SA-115228
Journal ID: ISSN 2326-5094; 400904120
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Resource Relation:
Journal Name: Health Security; Journal Volume: 15; Journal Issue: 1
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Ozanich, Richard M., Colburn, Heather A., Victry, Kristin D., Bartholomew, Rachel A., Arce, Jennifer S., Heredia-Langner, Alejandro, Jarman, Kristin, Kreuzer, Helen W., and Bruckner-Lea, Cynthia J. Evaluation of PCR Systems for Field Screening of Bacillus anthracis. United States: N. p., 2017. Web. doi:10.1089/hs.2016.0043.
Ozanich, Richard M., Colburn, Heather A., Victry, Kristin D., Bartholomew, Rachel A., Arce, Jennifer S., Heredia-Langner, Alejandro, Jarman, Kristin, Kreuzer, Helen W., & Bruckner-Lea, Cynthia J. Evaluation of PCR Systems for Field Screening of Bacillus anthracis. United States. doi:10.1089/hs.2016.0043.
Ozanich, Richard M., Colburn, Heather A., Victry, Kristin D., Bartholomew, Rachel A., Arce, Jennifer S., Heredia-Langner, Alejandro, Jarman, Kristin, Kreuzer, Helen W., and Bruckner-Lea, Cynthia J. Wed . "Evaluation of PCR Systems for Field Screening of Bacillus anthracis". United States. doi:10.1089/hs.2016.0043.
@article{osti_1344637,
title = {Evaluation of PCR Systems for Field Screening of Bacillus anthracis},
author = {Ozanich, Richard M. and Colburn, Heather A. and Victry, Kristin D. and Bartholomew, Rachel A. and Arce, Jennifer S. and Heredia-Langner, Alejandro and Jarman, Kristin and Kreuzer, Helen W. and Bruckner-Lea, Cynthia J.},
abstractNote = {There is little published data on the performance of hand-portable polymerase chain reaction (PCR) instruments that could be used by first responders to determine if a suspicious powder contains a potential biothreat agent. We evaluated five commercially available hand-portable PCR instruments for detection of Bacillus anthracis (Ba). We designed a cost-effective, statistically-based test plan that allows instruments to be evaluated at performance levels ranging from 0.85-0.95 lower confidence bound (LCB) on the probability of detection (POD) at confidence levels of 80-95%. We assessed specificity using purified genomic DNA from 13 Ba strains and 18 Bacillus near neighbors, interference with 22 common hoax powders encountered in the field, and PCR inhibition when Ba spores were spiked into these powders. Our results indicated that three of the five instruments achieved >0.95 LCB on the POD with 95% confidence at test concentrations of 2,000 genome equivalents/mL (comparable to 2,000 spores/mL), displaying more than sufficient sensitivity for screening suspicious powders. These instruments exhibited no false positive results or PCR inhibition with common hoax powders, and reliably detected Ba spores spiked into common hoax powders, though some issues with instrument controls were observed. Our testing approach enables efficient instrument performance testing to a statistically rigorous and cost-effective test plan to generate performance data that will allow users to make informed decisions regarding the purchase and use of biodetection equipment in the field.},
doi = {10.1089/hs.2016.0043},
journal = {Health Security},
number = 1,
volume = 15,
place = {United States},
year = {Wed Feb 01 00:00:00 EST 2017},
month = {Wed Feb 01 00:00:00 EST 2017}
}