RAZOR™ EX Anthrax Air Detection System for Detection of Bacillus anthracis Spores from Aerosol Collection Samples: Collaborative Study

Abstract

The RAZOR EX Anthrax Air Detection System was validated in a collaborative study for the detection of Bacillus anthracis in aerosol collection buffer. Phosphate-buffered saline was charged with 1 mg/mL standardized dust to simulate an authentic aerosol collection sample. The dust-charged buffer was spiked with either B. anthracis Ames at 2000 spores/mL or Bacillus cereus at 20 000 spores/mL. Twelve collaborators participated in the study, with four collaborators at each of three sites. Each collaborator tested 12 replicates of B. anthracis in dust-charged buffer and 12 replicates of B. cereus in dust-charged buffer. All samples sets were randomized and blind-coded. All collaborators produced valid data sets (no collaborators displayed systematic errors) and there was only one invalid data point. After unblinding, the analysis revealed a cross-collaborator probability of detection (CPOD) of 1.00 (144 positive results from 144 replicates, 95% confidence interval 0.975-1.00) for the B. anthracis samples and a CPOD of 0.00 (0 positive results from 143 replicates, 95% confidence interval 0.00-0.0262) for the B. cereus samples. These data meet the requirements of AOAC Standard Method Performance Requirement 2010.003, developed by the Stakeholder Panel on Agent Detection Assays.

Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification ofBacillus anthracisSpores in Suspicious White Powders and Environmental Samples

“…However, PCR requires clean samples in a small volume and is not generally suitable for field use. 42,43 Other methods of detection have included fluorescence-based sandwich immunoassays on glass slides, 44,45 peptide functionalized surface-enhanced Raman spectroscopy (SERS), piezo-electric based detection, 43,46 PCR combined with fluorescence resonance energy transfer (FRET), 47 and aptamers and bacteriophage. 43 While each of these methods holds some promise for laboratory-based detection, none is currently appropriate for field use to rapidly screen unknown environmental samples (ie, white powders) for the presence of B. anthracis spores.…”

Detection and Identification of Bacillus anthracis: From Conventional to Molecular Microbiology Methods

“…POCKIT enables analysis of up to eight samples at a time and for B. anthracis detection three assays targeting pXO1, pXO2, and chromosomal PL3 markers are available. The reagents are produced in a dry format, but recommended storage temperature is 4 • C. The reaction time is about one hour for all the systems [51][52][53][54].…”

Evaluation of microbial qPCR workflows using engineered Saccharomyces cerevisiae

“…However, there remains a scarcity of standards, reference materials, and third-party testing to demonstrate the reliability of these technologies in the hands of end users, despite considerable efforts by the stakeholder community and Federal government. Only a few commercially available biodetection technologies have been submitted to third-party validation, including the Razor™ EX BioDetection System [7] , a qPCR-based assay.…”