Two-Component Direct Fluorescent-Antibody Assay for Rapid Identification of<i>Bacillus anthracis</i>

De, Barun K.; Bragg, Sandra L.; Sanden, Gary N.; Wilson, Kathy E.; Diem, Lois; Marston, Chung K.; Hoffmaster, Alex R.; Barnett, Gwen A.; Weyant, Robbin S.; Abshire, Teresa G.; Ezzell, John W.; Popović, Tanja

doi:10.3201/eid0810.020392

Cited by 68 publications

(52 citation statements)

References 19 publications

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“…G9241 produced a positive reaction with mAb specific to the B. anthracis cell-wall antigen but not with Ab against the B. anthracis capsule. Because it is not uncommon for some clinical isolates of B. cereus to react with the cell-wall-specific Ab, the positive reaction with this Ab was not unusual (16). Although the isolate did not react with the B. anthracis-specific capsule Ab, it did produce a capsule that was expressed during growth on sheep blood agar plates or in horse serum (Fig.…”

Section: Isolation Of B Cereus G9241 During a Retrospective Analysimentioning

confidence: 99%

Identification of anthrax toxin genes in aBacillus cereusassociated with an illness resembling inhalation anthrax

Hoffmaster

Ravel

Rasko

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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537

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Bacillus anthracis is the etiologic agent of anthrax, an acute fatal disease among mammals. It was thought to differ from Bacillus cereus, an opportunistic pathogen and cause of food poisoning, by the presence of plasmids pXO1 and pXO2, which encode the lethal toxin complex and the poly-␥-D-glutamic acid capsule, respectively. This work describes a non-B. anthracis isolate that possesses the anthrax toxin genes and is capable of causing a severe inhalation anthrax-like illness. Although initial phenotypic and 16S rRNA analysis identified this isolate as B. cereus, the rapid generation and analysis of a high-coverage draft genome sequence revealed the presence of a circular plasmid, named pBCXO1, with 99.6% similarity with the B. anthracis toxin-encoding plasmid, pXO1. Although homologues of the pXO2 encoded capsule genes were not found, a polysaccharide capsule cluster is encoded on a second, previously unidentified plasmid, pBC218. A͞J mice challenged with B. cereus G9241 confirmed the virulence of this strain. These findings represent an example of how genomics could rapidly assist public health experts responding not only to clearly identified select agents but also to novel agents with similar pathogenic potentials. In this study, we combined a public health approach with genome analysis to provide insight into the correlation of phenotypic characteristics and their genetic basis.

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Section: Isolation Of B Cereus G9241 During a Retrospective Analysimentioning

confidence: 99%

Identification of anthrax toxin genes in aBacillus cereusassociated with an illness resembling inhalation anthrax

Hoffmaster

Ravel

Rasko

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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444

537

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“…Serology and reverse transcription (RT)-PCR were performed by other laboratories at CDC. [42][43][44] § Contained a small, superficial, shave biopsy that was inadequate for pathologic evaluation.…”

Section: Methodsmentioning

confidence: 99%

The Critical Role of Pathology in the Investigation of Bioterrorism-Related Cutaneous Anthrax

Shieh

Guarner

Paddock

et al. 2003

The American Journal of Pathology

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Cutaneous anthrax is a rare zoonotic disease in the United States. The clinical diagnosis traditionally has been established by conventional microbiological methods, such as culture and gram staining. However, these methods often yield negative results when patients have received antibiotics. During the bioterrorism event of 2001, we applied two novel immunohistochemical assays that can detect Bacillus anthracis antigens in skin biopsy samples even after prolonged antibiotic treatment. These assays provided a highly sensitive and specific method for the diagnosis of cutaneous anthrax, and were critical in the early and rapid diagnosis of 8 of 11 cases of cutaneous anthrax during the outbreak investigation. Skin biopsies were obtained from 10 of these 11 cases, and histopathological findings included various degrees of ulceration, hemorrhage, edema, coagulative necrosis, perivascular inflammation, and vasculitis. Serology was also an important investigation tool, but the results required several weeks because of the need to test paired serum specimens. Other tests, including culture, special stains, and polymerase chain reaction assay, were less valuable in the diagnosis and epidemiological investigation of these cutaneous anthrax cases. This report underscores the critical role of pathology in investigating potential bioterrorism events and in guiding epidemiological studies, Anthrax captured worldwide attention and aroused serious public health concerns following the intentional release of the etiological agent Bacillus anthracis in the United States postal system during Fall 2001, 1,2 resulting in 11 cases of inhalational anthrax and 11 cases of cutaneous anthrax. [3][4][5][6][7] In its conventional form, cutaneous anthrax accounts for 95% of all naturally occurring B. anthracis infections in the United States. 8 -11 Patients often have a history of occupational contact with animals or animal products contaminated with B. anthracis spores. 12-14 These pathogenic spores are introduced through a cutaneous cut or abrasion, with the most common areas of exposure are the head, neck, and extremities, although any area can be involved. Bacteremia and toxemia following cutaneous infection can occur with a fatality rate of 20% to 25% among untreated cases. [15][16][17] Cutaneous anthrax is characterized by the formation of a black eschar surrounded by prominent edema and vesicles, which may resemble many other skin lesions, such as the brown recluse spider bite, 18 -21 ulceroglandular tularemia, 22,23 plague, 24,25 ecthyma gangrenosum, 26,27 various spotted fever group rickettsial infections, 28 -31 and scrub typhus. 32,33 The clinical diagnosis of cutaneous anthrax is traditionally established by microbiological methods (eg, demonstrating gram-positive, capsulated bacilli on the smear of the lesion or isolating B. anthracis in culture). 34,35 However, gram stain and culture for B. anthracis can be unrevealing for patients who receive antibiotic therapy before specimens are obtained. 36,37 Historically, skin biopsies ...

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“…A number of further diagnostic tests may be employed to confirm IA cases; these fall broadly into assays which detect the presence of the bacteria, such as fluorescence immunoassays and mass spectrometry identifying cell-associated proteins [25][26][27] and assays which detect the presence of antibodies to PA and LF [28,29]. Several caveats must be borne in mind though when considering these methods, as phenotypic characteristics may vary between strains and hemolytic, motile bacilli, which are resistant to phage lysis, have been identified in B. anthracis isolates, closely related Bacillus species may lead to misdiagnosis, and (perhaps most importantly) antimicrobial therapy rapidly decreases the sensitivity of these assays [30].…”

Section: Anthrax Toxins and Disease Coursementioning

confidence: 99%

Anthrax in injecting drug users: the need for increased vigilance in the clinic

Ascough

Altmann

2015

Expert Review of Anti-infective Therapy

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Two-Component Direct Fluorescent-Antibody Assay for Rapid Identification ofBacillus anthracis

Cited by 68 publications

References 19 publications

Identification of anthrax toxin genes in aBacillus cereusassociated with an illness resembling inhalation anthrax

Identification of anthrax toxin genes in aBacillus cereusassociated with an illness resembling inhalation anthrax

The Critical Role of Pathology in the Investigation of Bioterrorism-Related Cutaneous Anthrax

Anthrax in injecting drug users: the need for increased vigilance in the clinic

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