Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification of<i>Bacillus anthracis</i>Spores in Suspicious White Powders and Environmental Samples

Ramage, J.; Prentice, Kristin; DePalma, Lindsay; Venkateswaran, K.; Chivukula, Sruti; Chapman, Carol; Bell, Melissa; Datta, Shomik; Singh, Ajay K.; Hoffmaster, Alex R.; Sarwar, Jawad; Parameswaran, Nishanth; Joshi, Mrinmayi; Thirunavkkarasu, Nagarajan; Krishnan, Viswanathan V; Morse, Stephen; Avila, Julie R.; Sharma, Shashi; Estacio, Peter L.; Stanker, Larry H.; Hodge, David R.; Pillai, Segaran P.

doi:10.1089/hs.2016.0041

Cited by 23 publications

(19 citation statements)

References 45 publications

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“…We, therefore, propose that the newly developed methodology described in this work allows the detection of spore concentrations that are 3-4 orders of magnitude lower than published immuno-affinity assays [11,36], being comparable to sensitive PCR-based assays [7,8,20], while being performed in under 3.5 h for the total assay. This time frame could potentially be shortened even further by modifying the multiplex assay step to be performed as a one-step assay (all in one) by using direct labeled secondary antibodies or a biotin-streptavidin complex for signal enhancement.…”

Section: Discussionmentioning

confidence: 98%

“…Additional identification assays have low sensitivities for environmental samples, suffering a 3-4 orders of magnitude decrease in sensitivity compared to pure samples [10]. This was demonstrated using specific phage [35] or other commercial assays [36]. The detection of B. anthracis spores from environmental samples in various immune affinity-based assays has been shown to be effective for the detection of 1 × 10 5 -1 × 10 6 spores [11,36].…”

Section: Discussionmentioning

confidence: 99%

“…This was demonstrated using specific phage [35] or other commercial assays [36]. The detection of B. anthracis spores from environmental samples in various immune affinity-based assays has been shown to be effective for the detection of 1 × 10 5 -1 × 10 6 spores [11,36]. In addition, using specific reporter phage enables the detection of 1 × 10 5 CFU in assays, taking several hours to completion [35].…”

Section: Discussionmentioning

confidence: 99%

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Rapid and Sensitive Multiplex Assay for the Detection of B. anthracis Spores from Environmental Samples

et al. 2020

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Prompt and accurate detection of Bacillus anthracis spores is crucial in the event of intentional spore dissemination in order to reduce the number of expected casualties. Specific identification of these spores from environmental samples is both challenging and time-consuming. This is due to the high homology with other Bacillus species as well as the complex composition of environmental samples, which further impedes assay sensitivity. Previously, we showed that a short incubation of B.anthracis spores in a defined growth medium results in rapid germination, bacterial growth, and secretion of toxins, including protective antigen. In this work, we tested whether coupling the incubation process to a newly developed immune-assay will enable the detection of secreted toxins as markers for the presence of spores in environmental samples. The new immune assay is a flow cytometry-based multiplex that simultaneously detects a protective antigen, lethal factor, and edema factor. Our combined assay detects 1 × 103–1 × 104/mL spores after a 2 h incubation followed by the ~80 min immune-multiplex detection. Extending the incubation step to 5 h increased assay sensitivity to 1 × 102/mL spore. The protocol was validated in various environmental samples using attenuated or fully virulent B. anthracis spores. There was no substantial influence of contaminants derived from real environmental samples on the performance of the assay compared to clean samples, which allow the unequivocal detection of 3 × 103/mL and 3 × 102/mL spores following 2 and 5 hour’s incubation, respectively. Overall, we propose this method as a rapid, sensitive, and specific procedure for the identification of B. anthracis spores in environmental samples.

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Rapid and Sensitive Multiplex Assay for the Detection of B. anthracis Spores from Environmental Samples

et al. 2020

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“…They were able to detect F. tularensis in both lemming carcasses and the well water in which the carcasses were found; however, this assay was less sensitive than PCR. Rapid BTA assays have previously been evaluated for the detection of biothreat agents including orthopoxviruses, 36 ricin, 37 abrin, 38 Bacillus anthracis, 32,39 and Yersinia pestis. 40 Limited evaluations have also been conducted with assays for the detection of Yersinia pestis, 41 botulinum neurotoxins, 42,43 and staphylococcal enterotoxins.…”

Section: Discussionmentioning

confidence: 99%

“…Table 3 shows the information about the 61 strains of diverse environmental background organisms used in the study. 32 Each of the microorganisms was inoculated onto optimal medium and incubated under appropriate conditions for 24 to 48 hours. A single, isolated colony was selected and inoculated onto a second agar plate and incubated for 1 to 6 days, depending on the organism and its growth rate.…”

Section: Phase 3: Near Neighbor Panelmentioning

confidence: 99%

Comprehensive Laboratory Evaluation of a Specific Lateral Flow Assay for the Presumptive Identification of Francisella tularensis in Suspicious White Powders and Aerosol Samples

et al. 2020

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We conducted a comprehensive, multi-phase laboratory evaluation of the Tularemia BioThreat Alert Ò (BTA) test, a lateral flow assay (LFA) for the rapid detection of Francisella tularensis. The study, conducted at 2 sites, evaluated the limit of detection (LOD) of this assay using the virulent SchuS4 strain and the avirulent LVS strain of F. tularensis. In 6-phase evaluation (linear dynamic range and reproducibility, inclusivity, near-neighbor, environmental background, white powder, and environmental filter extract), 13 diverse strains of F. tularensis, 8 Francisella near neighbors, 61 environmental background organisms, 26 white powders, and a pooled aerosol extract were tested. In the 937 tests performed, the Tularemia BTA demonstrated an LOD of 10 7 to 10 8 cfu/mL, with a sensitivity of 100.00%, specificity of 98.08%, and accuracy of 98.84%. These performance data are important for accurate interpretation of qualitative results arising from screening suspicious white powders in the field.

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Bacillus and Other Aerobic Endospore‐Forming Bacteria

Turenne,

Alexander

2023

ClinMicroNow

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Historically, most aerobic endospore‐forming bacteria were classified as species of Bacillus . Bacillus is the type genus of the Bacillaceae and contains the largest number of species. Aerobic endospore‐forming bacteria are ubiquitous in nature. Cases of anthrax due to laboratory accidents, biological warfare, and bioterrorism events are also rare, but ongoing vigilance in the clinical microbiology laboratory is critical. This chapter describes the clinically significant infections involving Bacillus , Brevibacillus , Lysinibacillus , and Paenibacillus species. Members of the B. cereus group, other than B. anthracis ,have been isolated from diverse ecological niches and are widely distributed in nature. Gastrointestinal anthrax is suspected only if an adequate history of the patient is known. Anthrax meningitis may occur on its own or, more frequently, as a complication of another form of the disease.

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

Cited by 23 publications

References 45 publications

Rapid and Sensitive Multiplex Assay for the Detection of B. anthracis Spores from Environmental Samples

Rapid and Sensitive Multiplex Assay for the Detection of B. anthracis Spores from Environmental Samples

Comprehensive Laboratory Evaluation of a Specific Lateral Flow Assay for the Presumptive Identification of Francisella tularensis in Suspicious White Powders and Aerosol Samples

Bacillus and Other Aerobic Endospore‐Forming Bacteria

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