Advances in Anthrax Detection: Overview of Bioprobes and Biosensors

Kim, Joungmok; Gedi, Vinayakumar; Lee, Sang-Choon; Cho, Jun‐Haeng; Moon, Ji‐Young; Yoon, Moon‐Young

doi:10.1007/s12010-015-1625-z

Cited by 40 publications

(24 citation statements)

References 133 publications

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“…cereus^group including B. anthraics, B. cereus, and B. thuringensis [5], these isolates, therefore, were indistinguished from B. cereus used as reference strains. Thus, the use of both plasmids pXO1 and pXO2 in combination to accurately identify pathogenic B. anthracis was recommended [13,30,31]. Here, the three pathogenic B. anthracis were explored by the hexaplex PCR assay which was to the best of our knowledge the first report for cases of anthrax during two past decades in Vietnam.…”

Section: Discussionmentioning

confidence: 99%

A Hexaplex PCR Assay Developed for Simultaneous Detection of Bacillus anthracis and Yersinia pestis and Distinguish Their Virulence Levels Tested on Vietnamese Samples

Nguyen

Dinh

Nghiêm

2019

SN Compr. Clin. Med.

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Bacillus anthracis and Yersinia pestis are at high risk of bioterrorism, but they have different strains of virulence that exist in the natural environment, making it difficult to diagnose the risk of disease. This study aimed to develop a novel hexaplex PCR assay to simultaneously detect B. anthracis and Y. pestis and quickly distinguish their virulence levels. The vrrA, pagA, and capA genes from B. anthracis and ypo2088, pla, and caf1 genes from Y. pestis were specifically amplified to generate six PCR fragments of 222, 751, and 610 and 103, 438, and 271 bp, respectively. Furthermore, a recombinant competitive internal amplification control was designed in the assay, which coamplified with the capA gene primer pairs, producing a fragment of 1000 bp in length. Thirtyeight bacterial strains including 4 reference strains and 34 screening strains derived from human, herbivores, and environments in the North of Vietnam, as well as spiked soil samples, were used to evaluate the assay. The new hexaplex PCR assay was in accordance with the conventional culture methods to detect B. anthracis and Y. pestis. In addition, this assay can be quickly discriminated among pathogenic and non-pathogenic B. anthracis and Y. pestis strains, as well as rapidly differentiate levels of virulence among strains isolated from human and soil samples in some areas of Vietnam. The new hexaplex PCR is a useful tool in screening for both pathogenic and non-pathogenic B. anthracis and Y. pestis, necessary in use for biodefense purposes and disease prevention.

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Section: Discussionmentioning

confidence: 99%

A Hexaplex PCR Assay Developed for Simultaneous Detection of Bacillus anthracis and Yersinia pestis and Distinguish Their Virulence Levels Tested on Vietnamese Samples

Nguyen

Dinh

Nghiêm

2019

SN Compr. Clin. Med.

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“…Conventional cultivation methods are clearly inappropriate under such conditions, because they depend on highly skilled experts, require specific biosafety level 3 (BSL 3) facilities, and have turnaround times of several hours or days ( 8 ). The high specificity, sensitivity, and speed of nucleic acid molecular methods, including real-time quantitative PCR (qPCR), explain why these methods have become a predominant diagnostic tool ( 9 , 10 ). In addition, novel DNA-based technologies based on isothermal amplification are now steadily gaining interest among first responders, because of their simplicity, speed, and appropriateness for in-field use.…”

Section: Introductionmentioning

confidence: 99%

“…However, detecting B. anthracis chromosomal signature sequences has proved to be very challenging. Among all reported targets, only a few, e.g., purA and pclR single-nucleotide polymorphisms (SNPs), as well as BA_5345, PL3 , and BA5357 genes, were found to be unique to B. anthracis ( 9 , 25 ).…”

Section: Introductionmentioning

confidence: 99%

Sensitive and Specific Recombinase Polymerase Amplification Assays for Fast Screening, Detection, and Identification of Bacillus anthracis in a Field Setting

Bentahir

Ambroise

Delcorps³

et al. 2018

Appl Environ Microbiol

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Four isothermal recombinase polymerase amplification (RPA) assays were developed for fast in-field identification of Bacillus anthracis. The RPA assays targeted three specific sequences (i.e., the BA_5345 chromosomal marker, the lethal factor lef [from pXO1], and the capsule-biosynthesis-related capA [from pXO2]) and a conserved sequence in the adenylate cyclase gene (adk) for the Bacillus cereus group. B. anthracis-specific RPA assays were tested first with purified genomic DNAs (n = 60), including 11 representatives of B. anthracis, and then with soil (n = 8) and white powder (n = 8) samples spiked with inactivated B. anthracis spores and/or other biological agents. The RPA assays were also tested in another laboratory facility, which blindly provided DNA and lysate samples (n = 30, including 20 B. anthracis strains). RPA assays displayed 100% specificity and sensitivity. The hands-off turnaround times at 42°C ranged from 5 to 6 min for 102 genomic copies. The analytical sensitivity of each RPA assay was ∼10 molecules per reaction. In addition, the BA_5345 and adk RPA assays were assessed under field conditions with a series of surface swabs (n = 13, including 11 swabs contaminated with B. thuringiensis spores) that were blindly brought to the field laboratory by a chemical, biological, radiological, and nuclear (CBRN) sampling team. None of the 13 samples, except the control, tested positive for B. anthracis, and all samples that had been harvested from spore-contaminated surfaces tested positive with the adk RPA assay. All three B. anthracis-specific RPA assays proved suitable for rapid and reliable identification of B. anthracis and therefore could easily be used by first responders under field conditions to quickly discriminate between a deliberate release of B. anthracis spores and a hoax attack involving white powder.IMPORTANCE In recent decades, particularly following the 11 September 2001 and Amerithrax attacks, the world has experienced attempts to sow panic and chaos in society through thousands of white-powder copycats using household powders to mimic real bioterrorism attacks. In such circumstances, field-deployable detection methods are particularly needed to screen samples collected from the scene. The aim is to test the samples directly using a fast and reliable assay for detection of the presence of B. anthracis. While this would not preclude further confirmatory tests from being performed in reference laboratories, it would bring useful, timely, and relevant information to local crisis managers and help them make appropriate decisions without having to wait for quantitative PCR results (with turnaround times of a few hours) or phenotypic identification and sequencing (with turnaround times of a few days). In the current investigation, we developed a set of isothermal RPA assays for the rapid screening and identification of B. anthracis in powders and soil samples, with the purpose of discriminating a deliberate release of B. anthracis spores from a hoax attack involving white powder; th...

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“…There are a number of markers of inhalation anthrax infection including PA, LF and PGA; however, it has been shown that LF is detectable earliest in the infection [ 10 ]. An overview of anthrax detection methods has been published recently [ 11 ]. Highly sensitive, activity-based assays to detect LF in serum and plasma have been reported using either matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) or electrospray ionization MS/MS [ 12 – 15 ].…”

Section: Introductionmentioning

confidence: 99%

Design and use of a novel substrate for simple, rapid, and specific early detection of anthrax infection

Suryadi

Shine

2018

PLoS ONE

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Bacillus anthracis is a major biological warfare threat. The inhalation form of infection can kill quickly. While antibiotic treatment is effective, if diagnosis is delayed, the rapidly produced toxin may already be present in lethal amounts. This report describes a fast, sensitive, specific and accurate method for detection of active infection by Bacillus anthracis in plasma. One of the virulence factors, anthrax lethal factor, is an endopeptidase present in blood early in the infection. However, the use of peptidic substrates to detect endopetidases is problematic in plasma due to the presence of other proteases and the likelihood of nonspecific cleavage of the substrate. The fluorescently labeled peptide substrate MAPKKide Plus designed in this study is not cleaved by plasma proteases and thus is specific for lethal factor. Three detection strategies are described. Two include enrichment by capture from plasma using lethal factor antibody-coated microtiter plates or similarly coated immuno-tubes. The captured lethal factor is exposed to the MAPKKide Plus, and the amount of cleavage is determined either by HPLC or microplate reader. Concentration of lethal factor using the antibody-coated plates aplnd HPLC allows for detection of less than 5 pg lethal factor/ml of neat plasma after 2 hours of incubation. Using antibody-coated immuno-tubes, 20 pg lethal factor/ml plasma can be detected in 5 hours by a simple end point read of fluorescence in a microplate reader. For a third strategy, the substrate is added directly to diluted plasma, and cleavage is monitored by the increase in fluorescence as a function of time. The limit of detection by this simple method is 25 ng lethal factor/ml of plasma in 15 minutes, 5 ng/ml after 45 minutes, and <1 ng lethal factor/ml of plasma after 5 hours.

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Advances in Anthrax Detection: Overview of Bioprobes and Biosensors

Cited by 40 publications

References 133 publications

A Hexaplex PCR Assay Developed for Simultaneous Detection of Bacillus anthracis and Yersinia pestis and Distinguish Their Virulence Levels Tested on Vietnamese Samples

A Hexaplex PCR Assay Developed for Simultaneous Detection of Bacillus anthracis and Yersinia pestis and Distinguish Their Virulence Levels Tested on Vietnamese Samples

Sensitive and Specific Recombinase Polymerase Amplification Assays for Fast Screening, Detection, and Identification of Bacillus anthracis in a Field Setting

Design and use of a novel substrate for simple, rapid, and specific early detection of anthrax infection

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