Comparison of quantitative PCR and culture-based methods for evaluating dispersal of Bacillus thuringiensis endospores at a bioterrorism hoax crime scene

Crighton, Taryn; Hoile, Rebecca Jane; Coleman, Nicholas V.

doi:10.1016/j.forsciint.2011.12.003

Cited by 8 publications

(6 citation statements)

References 44 publications

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“…Historically, B. anthracis has played a central role as a biological warfare agent, but in 2001, the mailing of letters containing powders of B. anthracis spores to news media and government offices in the United States (the Amerithrax incident) killed five people, sickened an additional 17, and resulted in economic disbursements of more than one billion U.S. dollars, demonstrating B. anthracis as a bioterrorism agent (3,4). The majority of incidents involving suspicious powders eventually turn out to be hoaxes (i.e., the powder does not contain B. anthracis spores) (5). However, even hoax incidents have economic and psychological impacts, since they cannot easily be discriminated from bioterrorism incidents and must therefore be handled as real threats until the presence of B. anthracis spores or other hazards can be ruled out.…”

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confidence: 99%

Rapid Identification of Bacillus anthracis Spores in Suspicious Powder Samples by Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)

Dybwad

Laaken²,

Blatny

et al. 2013

Appl Environ Microbiol

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cRapid and reliable identification of Bacillus anthracis spores in suspicious powders is important to mitigate the safety risks and economic burdens associated with such incidents. The aim of this study was to develop and validate a rapid and reliable laboratory-based matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis method for identifying B. anthracis spores in suspicious powder samples. A reference library containing 22 different Bacillus sp. strains or hoax materials was constructed and coupled with a novel classification algorithm and standardized processing protocol for various powder samples. The method's limit of B. anthracis detection was determined to be 2.5 ؋ 10 6 spores, equivalent to a 55-g sample size of the crudest B. anthracis-containing powder discovered during the 2001 Amerithrax incidents. The end-to-end analysis method was able to successfully discriminate among samples containing B. anthracis spores, closely related Bacillus sp. spores, and commonly encountered hoax materials. No false-positive or -negative classifications of B. anthracis spores were observed, even when the analysis method was challenged with a wide range of other bacterial agents. The robustness of the method was demonstrated by analyzing samples (i) at an external facility using a different MALDI-TOF MS instrument, (ii) using an untrained operator, and (iii) using mixtures of Bacillus sp. spores and hoax materials. Taken together, the observed performance of the analysis method developed demonstrates its potential applicability as a rapid, specific, sensitive, robust, and cost-effective laboratory-based analysis tool for resolving incidents involving suspicious powders in less than 30 min.

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confidence: 99%

Rapid Identification of Bacillus anthracis Spores in Suspicious Powder Samples by Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)

Dybwad

Laaken²,

Blatny

et al. 2013

Appl Environ Microbiol

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“…If this test was used for diagnosis, laboratory tests with other methods must also be carried out. [1,7,18,19] The choice of method for laboratory examination was based on the target detected.…”

Section: Discussionmentioning

confidence: 99%

Laboratory Conﬁrmation Human Specimens Suspected of Anthrax in Kulonprogo District of Yogyakarta Special Region

Khariri

Muna

Rukminiati

2019

KSS

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Anthrax is an acute zoonoses infectious disease caused by Bacillus anthracis bacteria. Bacillus anthracis had ability to form endospores for self-defense. Anthrax infectioncouldbedividedintofourtypesnamelyskinanthrax,gastrointestinalanthrax, anthrax of the respiratory tract (lung) and meningitis anthrax. The examination was performed to conﬁrm the diagnosis of patients suspected of being infected with Bacillus anthracis. The examination methods consisted of culture for whole blood spesimens, konvensional polymerase chain reaction (PCR) on growing colonies and whole blood specimens without growing colony, and enzyme-linked immunosorbent assay (ELISA) for blood serum spesimens. The 6 cultured specimens, there were 2 colony-growing specimens with 1 identical colony with Bacillus anthracis colonies. The colonies conﬁrmed with PCR and only 1 positive as Bacillus anthracis pathogens. The 6 specimens examined there was a positive specimen of Bacillus anthracis pathogen.

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“…As indicated earlier, security and defense-related applications overlap considerably with other fields of use, as chemical and biological agents can be spread by multiple modes of dissemination (food, air, water), and therefore myriad sample types may be encountered; similarly, sample types used for drug testing can also vary from purified stocks to environmental samples to clinical fluids, depending on the intended purpose. Furthermore, the effects of other interfering materials – potentially used to mimic (Crighton et al 2012) or mask (Krebs et al 2000) the presence of target – must be accounted for.…”

Section: 0 Applications Of Evanescent Wave Fluorescence Biosensorsmentioning

confidence: 99%

Evanescent wave fluorescence biosensors: Advances of the last decade

Taitt

Anderson

Ligler

2016

Biosensors and Bioelectronics

125

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Biosensor development has been a highly dynamic field of research and has progressed rapidly over the past two decades. The advances have accompanied the breakthroughs in molecular biology, nanomaterial sciences, and most importantly computers and electronics. The subfield of evanescent wave fluorescence biosensors has also matured dramatically during this time. Fundamentally, this review builds on our earlier 2005 review. While a brief mention of seminal early work will be included, this current review will focus on new technological developments as well as technology commercialized in just the last decade. Evanescent wave biosensors have found a wide array applications ranging from clinical diagnostics to biodefense to food testing; advances in those applications and more are described herein.

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Comparison of quantitative PCR and culture-based methods for evaluating dispersal of Bacillus thuringiensis endospores at a bioterrorism hoax crime scene

Cited by 8 publications

References 44 publications

Rapid Identification of Bacillus anthracis Spores in Suspicious Powder Samples by Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)

Rapid Identification of Bacillus anthracis Spores in Suspicious Powder Samples by Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)

Laboratory Conﬁrmation Human Specimens Suspected of Anthrax in Kulonprogo District of Yogyakarta Special Region

Evanescent wave fluorescence biosensors: Advances of the last decade

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