scholarly journals Fast and Sensitive Detection of Bacillus anthracis Spores by Immunoassay

2012 ◽  
Vol 78 (18) ◽  
pp. 6491-6498 ◽  
Author(s):  
Nathalie Morel ◽  
Hervé Volland ◽  
Julie Dano ◽  
Patricia Lamourette ◽  
Patricia Sylvestre ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Electrical Current ◽  
Biological Weapons ◽  
Meso Scale Discovery ◽  
Simple Method ◽  
Content Type ◽  
Bacillus Anthracis Spores ◽  
Luminescent Signal ◽  
Sample Purification ◽  
Phosphate Buffered Saline

ABSTRACTBacillus anthracisis one of the most dangerous potential biological weapons, and it is essential to develop a rapid and simple method to detectB. anthracisspores in environmental samples. The immunoassay is a rapid and easy-to-use method for the detection ofB. anthracisby means of antibodies directed against surface spore antigens. With this objective in view, we have produced a panel of monoclonal antibodies againstB. anthracisand developed colorimetric and electrochemiluminescence (ECL) immunoassays. Using Meso Scale Discovery ECL technology, which is based on electrochemiluminescence (ECL) detection utilizing a sulfo-Tag label that emits light upon electrochemical stimulation (using a dedicated ECL plate reader, an electrical current is placed across the microplate with electrodes integrated into the bottom of the plate, resulting in a series of electrically induced reactions leading to a luminescent signal), a detection limit ranging between 0.3 × 103and 103CFU/ml (i.e., 30 to 100 spores per test), depending on theB. anthracisstrain assayed, was achieved. In complex matrices (5 mg/ml of soil or simulated powder), the detection level (without any sample purification or concentration) was never altered more than 3-fold compared with the results obtained in phosphate-buffered saline.

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

2013 ◽  
Vol 96 (2) ◽  
pp. 392-398 ◽  
Author(s):  
Ted Hadfield ◽  
Valorie Ryan ◽  
Usha K Spaulding ◽  
Kristine M Clemens ◽  
Irene M Ota ◽  
...  
Keyword(s):  
Confidence Interval ◽  
Bacillus Anthracis ◽  
Detection System ◽  
Collaborative Study ◽  
Probability Of Detection ◽  
Data Sets ◽  
Method Performance ◽  
Bacillus Anthracis Spores ◽  
Phosphate Buffered Saline ◽  
Positive Results

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.

scholarly journals Evaluation of the Efficacy of Methyl Bromide in the Decontamination of Building and Interior Materials Contaminated with Bacillus anthracis Spores

2016 ◽  
Vol 82 (7) ◽  
pp. 2003-2011 ◽  
Author(s):  
Joseph P. Wood ◽  
Morgan Wendling ◽  
William Richter ◽  
Andrew Lastivka ◽  
Leroy Mickelsen
Keyword(s):  
Bacillus Anthracis ◽  
Methyl Bromide ◽  
Contact Time ◽  
Geobacillus Stearothermophilus ◽  
Efficacy Data ◽  
Content Type ◽  
New Information ◽  
Bacillus Anthracis Spores ◽  
Effective Use ◽  
Selection Of

ABSTRACTThe primary goal of this study was to determine the conditions required for the effective inactivation ofBacillus anthracisspores on materials by using methyl bromide (MeBr) gas. Another objective was to obtain comparative decontamination efficacy data with three avirulent microorganisms to assess their potential for use as surrogates forB. anthracisAmes. Decontamination tests were conducted with spores ofB. anthracisAmes andGeobacillus stearothermophilus,B. anthracisNNR1Δ1, andB. anthracisSterne inoculated onto six different materials. Experimental variables included temperature, relative humidity (RH), MeBr concentration, and contact time. MeBr was found to be an effective decontaminant under a number of conditions. This study highlights the important role that RH has when fumigation is performed with MeBr. There were no tests in which a ≥6-log10reduction (LR) ofB. anthracisAmes was achieved on all materials when fumigation was done at 45% RH. At 75% RH, an increase in the temperature, the MeBr concentration, or contact time generally improved the efficacy of fumigation with MeBr. This study provides new information for the effective use of MeBr at temperatures and RH levels lower than those that have been recommended previously. The study also provides data to assist with the selection of an avirulent surrogate forB. anthracisAmes spores when additional tests with MeBr are conducted.

scholarly journals Detection of Bacillus anthracis Spores Using Peptide Functionalized SERS-Active Substrates

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Atanu Sengupta ◽  
Chetan Shende ◽  
Stuart Farquharson ◽  
Frank Inscore
Keyword(s):  
Bacillus Anthracis ◽  
Dipicolinic Acid ◽  
Surface Enhanced Raman Spectroscopy ◽  
Biological Weapons ◽  
Low Concentrations ◽  
Biological Warfare Agents ◽  
Active Metal ◽  
Bacillus Anthracis Spores ◽  
Surface Enhanced Raman ◽  
Warfare Agents

The need for portable technologies that can rapidly identify biological warfare agents (BWAs) in the field remains an international priority as expressed at the 2011 Biological Weapons Convention. In recent years, the ability of surface-enhanced Raman spectroscopy (SERS) to rapidly detect various BWAs at very low concentrations has been demonstrated. However, in the specific case of Bacillus anthracis, differentiation at the species level is required since other bacilli are common in the environment, representing potential false-positive responses. To overcome this limitation, we describe the use of a peptide attached to the SERS-active metal that selectively binds Bacillus anthracis-Sterne as the target analyte. Using this approach, 109  B. anthracis-Sterne spores/mL produced an intense dipicolinic acid spectrum upon the addition of acetic acid, while the same concentration and treatment of B. cereus and B. subtilis did not.

scholarly journals Anthrax: How it kills, and how it can be stopped

2004 ◽  
Vol 26 (2) ◽  
pp. 15-17
Author(s):  
Jeremy Mogridge
Keyword(s):  
Bacillus Anthracis ◽  
Biological Weapon ◽  
Biological Weapons ◽  
The Public ◽  
Medical Researchers ◽  
Bacillus Anthracis Spores ◽  
The Usa ◽  
The Impact

The intentional release of Bacillus anthracis spores in the USA during the autumn of 2001 alerted the public to the possibility of future attacks using biological weapons. It also underscored the importance of understanding the pathogenesis of the organism and the need for developing new therapeutics and vaccines. Fortunately, there has been a resurgence in anthrax research. This review focuses on the characteristics of B. anthracis that have led to its use as a biological weapon and the approaches that are being taken by medical researchers to minimize the impact of another release.

scholarly journals Aerosol and Surface Deposition Characteristics of Two Surrogates for Bacillus anthracis Spores

2016 ◽  
Vol 82 (22) ◽  
pp. 6682-6690 ◽  
Author(s):  
Alistair H. Bishop ◽  
Helen L. Stapleton
Keyword(s):  
Bacillus Anthracis ◽  
Pathogenic Bacteria ◽  
Human Infection ◽  
Solid Surfaces ◽  
Surface Deposition ◽  
Content Type ◽  
Hazard Management ◽  
Bacillus Anthracis Spores ◽  
Air Blower ◽  
Set Up

ABSTRACTSpores of an acrystalliferous derivative ofBacillus thuringiensissubsp.kurstaki, termedBtcry−, are morphologically, aerodynamically, and structurally indistinguishable fromBacillus anthracisspores.Btcry− spores were dispersed in a large, open-ended barn together with spores ofBacillus atrophaeussubsp.globigii, a historically used surrogate forBacillus anthracis. Spore suspensions (2 × 1012CFU each ofB. atrophaeussubsp.globigiiandBtcry−) were aerosolized in each of five spray events using a backpack misting device incorporating an air blower; a wind of 4.9 to 7.6 m s−1was also flowing through the barn in the same direction. Filter air samplers were situated throughout the barn to assess the aerosol density of the spores during each release. Trays filled with a surfactant in aqueous buffer were placed on the floor near the filter samplers to assess spore deposition. Spores were also recovered from arrays of solid surfaces (concrete, aluminum, and plywood) that had been laid on the floor and set up as a wall at the end of the barn.B. atrophaeussubsp.globigiispores were found to remain airborne for significantly longer periods, and to be deposited on horizontal surfaces at lower densities, thanBtcry− spores, particularly near the spray source. There was a 6-fold-higher deposition ofBtcry− spores than ofB. atrophaeussubsp.globigiispores on vertical surfaces relative to the surrounding airborne density. This work is relevant for selecting the bestB. anthracissurrogate for the prediction of human exposure, hazard assessment, and hazard management following a malicious release ofB. anthracis.IMPORTANCEThere is concern that pathogenic bacteria could be maliciously disseminated in the air to cause human infection and disruption of normal life. The threat from spore-forming organisms, such as the causative agent of anthrax, is particularly serious. In order to assess the extent of this risk, it is important to have a surrogate organism that can be used to replicate the dispersal characteristics of the threat agent accurately. This work compares the aerosol dispersal and deposition behaviors of the surrogatesBtcry− andB. atrophaeussubsp.globigii.Btcry− spores remained in the air for a shorter time, and were markedly more likely to adhere to vertical surfaces, thanB. atrophaeussubsp.globigiispores.

scholarly journals Existence of Separate Domains in Lysin PlyG for Recognizing Bacillus anthracis Spores and Vegetative Cells

2012 ◽  
Vol 56 (10) ◽  
pp. 5031-5039 ◽  
Author(s):  
Hang Yang ◽  
Dian-Bing Wang ◽  
Qiuhua Dong ◽  
Zhiping Zhang ◽  
Zongqiang Cui ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Catalytic Domain ◽  
Vegetative Cells ◽  
Content Type ◽  
Bacillus Anthracis Spores ◽  
Acid Domain ◽  
New Biomarkers ◽  
First Time ◽  
Amino Acid Domain ◽  
Spore Forming Bacteria

ABSTRACTAs a potential antimicrobial, the bacteriophage lysin PlyG has been reported to specifically recognizeBacillus anthracisvegetative cells only and to killB. anthracisvegetative cells and its germinating spores. However, how PlyG interacts withB. anthracisspores remains unclear. Herein, a 60-amino-acid domain in PlyG (residues 106 to 165), located mainly in the previously identified catalytic domain, was found able to specifically recognizeB. anthracisspores but not vegetative cells. The exosporium of the spores was found to be the most probable binding target of this domain. This is the first time that a lysin for spore-forming bacteria has been found to have separate domains to recognize spores and vegetative cells, which might help in understanding the coevolution of phages with spore-forming bacteria. Besides providing new biomarkers for developing better assays for identifyingB. anthracisspores, the newly found domain may be helpful in developing PlyG as a preventive antibiotic to reduce the threat of anthrax in suspected exposures toB. anthracisspores.

scholarly journals Microbicidal Power of Alpha Radiation in Sterilizing Germinating Bacillus anthracis Spores

2013 ◽  
Vol 58 (3) ◽  
pp. 1813-1815 ◽  
Author(s):  
Johanna Rivera ◽  
Alfred Morgenstern ◽  
Frank Bruchertseifer ◽  
John F. Kearney ◽  
Charles L. Turnbough ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Dependent Manner ◽  
Alpha Radiation ◽  
Content Type ◽  
Immunodominant Antigen ◽  
Spore Killing ◽  
Bacillus Anthracis Spores ◽  
A Site ◽  
Antigen Antibody ◽  
Dose Dependent

ABSTRACTRadioimmunotherapy (RIT) takes advantage of the specificity and affinity of the antigen-antibody interaction to deliver microbicidal radioactive nuclides to a site of infection. In this study, we investigated the microbicidal properties of an alpha particle-emitting213Bi-labeled monoclonal antibody (MAb), EA2-1 (213Bi-EA2-1), that binds to the immunodominant antigen onBacillus anthracisspores. Our results showed that dormant spores were resistant to213Bi-EA2-1. Significant spore killing was observed following treatment with EA2-1 labeled with 300 μCi213Bi; however, this effect was not dependent on the MAb. In contrast, when spores were germinating,213Bi-EA2-1 mediated MAb-specific killing in a dose-dependent manner. Dormant spores are very resistant to RIT, and RIT should focus on targeting vegetative cells and germinating spores.

scholarly journals Nanomechanical Characterization of Bacillus anthracis Spores by Atomic Force Microscopy

2016 ◽  
Vol 82 (10) ◽  
pp. 2988-2999 ◽  
Author(s):  
Alex G. Li ◽  
Larry W. Burggraf ◽  
Yun Xing
Keyword(s):  
Atomic Force Microscopy ◽  
Bacillus Anthracis ◽  
Elevated Temperatures ◽  
Adhesion Force ◽  
Spore Surface ◽  
Content Type ◽  
Force Microscopy ◽  
Nanomechanical Properties ◽  
Atomic Force ◽  
Bacillus Anthracis Spores

ABSTRACTThe study of structures and properties of bacterial spores is important to understanding spore formation and biological responses to environmental stresses. While significant progress has been made over the years in elucidating the multilayer architecture of spores, the mechanical properties of the spore interior are not known. Here, we present a thermal atomic force microscopy (AFM) study of the nanomechanical properties of internal structures ofBacillus anthracisspores. We developed a nanosurgical sectioning method in which a stiff diamond AFM tip was used to cut an individual spore, exposing its internal structure, and a soft AFM tip was used to image and characterize the spore interior on the nanometer scale. We observed that the elastic modulus and adhesion force, including their thermal responses at elevated temperatures, varied significantly in different regions of the spore section. Our AFM images indicated that the peptidoglycan (PG) cortex ofBacillus anthracisspores consisted of rod-like nanometer-sized structures that are oriented in the direction perpendicular to the spore surface. Our findings may shed light on the spore architecture and properties.IMPORTANCEA nanosurgical AFM method was developed that can be used to probe the structure and properties of the spore interior. The previously unknown ultrastructure of the PG cortex ofBacillus anthracisspores was observed to consist of nanometer-sized rod-like structures that are oriented in the direction perpendicular to the spore surface. The variations in the nanomechanical properties of the spore section were largely correlated with its chemical composition. Different components of the spore materials showed different thermal responses at elevated temperatures.

scholarly journals Tetrazole-Based trans-Translation Inhibitors Kill Bacillus anthracis Spores To Protect Host Cells

2017 ◽  
Vol 61 (10) ◽  
Author(s):  
John N. Alumasa ◽  
Tyler D. P. Goralski ◽  
Kenneth C. Keiler
Keyword(s):  
Bacillus Anthracis ◽  
Host Cells ◽  
Vegetative Cells ◽  
Content Type ◽  
Chemical Agents ◽  
Germination Process ◽  
Bacillus Anthracis Spores ◽  
Lethal Doses ◽  
Potential Use

ABSTRACT Bacillus anthracis, the causative agent of anthrax, remains a significant threat to humans, including potential use in bioterrorism and biowarfare. The capacity to engineer strains with increased pathogenicity coupled with the ease of disseminating lethal doses of B. anthracis spores makes it necessary to identify chemical agents that target and kill spores. Here, we demonstrate that a tetrazole-based trans-translation inhibitor, KKL-55, is bactericidal against vegetative cells of B. anthracis in culture. Using a fluorescent analog, we show that this class of compounds colocalizes with developing endospores and bind purified spores in vitro. KKL-55 was effective against spores at concentrations close to its MIC for vegetative cells. Spore germination was inhibited at 1.2× MIC, and spores were killed at 2× MIC. In contrast, ciprofloxacin killed germinants at concentrations close to its MIC but did not prevent germination even at 32× MIC. Because toxins are released by germinants, macrophages infected by B. anthracis spores are killed early in the germination process. At ≥2× MIC, KKL-55 protected macrophages from death after infection with B. anthracis spores. Ciprofloxacin required concentrations of ≥8× MIC to exhibit a similar effect. Taken together, these data indicate that KKL-55 and related tetrazoles are good lead candidates for therapeutics targeting B. anthracis spores and suggest that there is an early requirement for trans-translation in germinating spores.

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