Chapter 63 : Detection and Anthrax Diagnosis

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:
Zoom in

Detection and Anthrax Diagnosis, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555815905/9781555813642_Chap63-1.gif /docserver/preview/fulltext/10.1128/9781555815905/9781555813642_Chap63-2.gif


This chapter focuses on methods that are currently in use within Laboratory Response Network (LRN) laboratories and on commercially available tests that have been approved by the U.S. Food and Drug Administration (FDA) and thus are widely available. The chapter includes four sections, covering culture, molecular, antigen, and antibody detection methods for the diagnosis of anthrax. Supportive laboratory tests including the LRN PCR assay, immunohistochemical (IHC) staining of tissues, and anti-protective antigen (anti-PA) immunoglobulin G (IgG) detection by an enzyme-linked immunosorbent assay (ELISA) are discussed in this chapter. LRN real-time PCR assay was also used during the 2001 bioterrorism-associated anthrax outbreak to detect in environmental samples and clinical specimens to confirm anthrax diagnosis when the isolation of failed due to the initiation of antimicrobial drug treatment. Antibodies have been used successfully to detect the vegetative and spore forms of . A section covers the following four methods: a direct fluorescent-antibody assay (DFA), time-resolved fluorescence (TRF) assays, an immunochromatographic assay, and IHC assays. In addition, quantitative ELISAs and the toxin neutralization assay (TNA) have also been used in vaccine-animal challenge studies to show that levels of anti-PA antibodies are a significant predictor of survival. Recently, a commercially available test was FDA approved to qualitatively measure anti-PA antibodies in human serum (QuickELISA Anthrax-PA kit). Current research interests and funding will continue to yield novel technologies and new methods for the detection of and the diagnosis of anthrax in the future.

Citation: Hoffmaster A, Popovic T. 2006. Detection and Anthrax Diagnosis, p 547-552. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch63

Key Concept Ranking

Anthrax Lethal Toxin
Enzyme-Linked Immunosorbent Assay
Highlighted Text: Show | Hide
Loading full text...

Full text loading...


1. Anonymous. 1999. Bacterial agents, p. 8889. In J. Y. Richmond and , R. W. McKinney (ed.), Biosafety in Microbiological and Biomedical Laboratories, 4th ed. U.S. Government Printing Office, Washington D.C.
2. Bell, C. A.,, J. R. Uhl,, T. L. Hadfield,, J. C. David,, R. F. Meyer,, T. F. Smith, and , F. R. Cockerill, III. 2002. Detection of Bacillus anthracis DNA by LightCycler PCR. J. Clin. Microbiol. 40:28972902.
3. Brown, E. R., and , W. B. Cherry. 1955. Specific identification of Bacillus anthracis by means of a variant bacterio-phage. J. Infect. Dis. 96:3439.
4. Carucci, J. A.,, T. W. McGovern,, S. A. Norton,, C. R. Daniel,, B. E. Elewski,, S. Fallon-Friedlander,, B. D. Lushniak,, J. S. Taylor,, K. Warschaw, and , R. G. Wheeland. 2002. Cutaneous anthrax management algorithm. J. Am. Acad. Dermatol. 47:766769.
5. Centers for Disease Control and Prevention. 2001. Update: Investigation of anthrax associated with intentional exposure and interim public health guidelines, October 2001. Morb. Mortal. Wkly. Rep. 50:889893.
6. Cockerill, F. R., and , T. F. Smith. 2002. Rapid-cycle realtime PCR: a revolution for clinical microbiology. ASM News 68:7783.
7. De, B. K.,, S. L. Bragg,, G. N. Sanden,, K. E. Wilson,, L. A. Diem,, C. K. Marston,, A. R. Hoffmaster,, G. A. Barnett,, R. S. Weyant,, T. G. Abshire,, J. W. Ezzell, and , T. Popovic. 2002. Two-component direct fluorescent-antibody assay for rapid identification of Bacillus anthracis. Emerg. Infect. Dis. 8:10601065.
8. Dragon, D. C.,, R. P. Rennie, and , B. T. Elkin. 2001. Detection of anthrax spores in endemic regions of northern Canada. J. Appl. Microbiol. 91:435441.
9. Ezzell, J. W., Jr., and , T. G. Abshire. 1988. Immunological analysis of cell-associated antigens of Bacillus anthracis. Infect. Immun. 56:349356.
10. Ezzell, J. W., Jr.,, T. G. Abshire,, S. F. Little,, B. C. Lidgerding, and , C. Brown. 1990. Identification of Bacillus anthracis by using monoclonal antibody to cell wall galactose-N-acetylglucosamine polysaccharide. J. Clin. Microbiol. 28:223231.
11. Guarner, J.,, J. A. Jernigan,, W. J. Shieh,, K. Tatti,, L. M. Flannagan,, D. S. Stephens,, T. Popovic,, D. A. Ashford,, B. A. Perkins, and , S. R. Zaki. 2003. Pathology and patho-genesis of bioterrorism-related inhalational anthrax. Am. J. Pathol. 163:701709.
12. Helgason, E.,, O. A. Okstad,, D. A. Caugant,, H. A. Johansen,, A. Fouet,, M. Mock,, I. Hegna, and , A.-B. Kolsto. 2000. Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis—one species on the basis of genetic evidence. Appl. Environ. Microbiol. 66:26272630.
13. Hering, D.,, W. Thompson,, J. Hewetson,, S. Little,, S. Norris, and , J. Pace-Templeton. 2004. Validation of the anthrax lethal toxin neutralization assay. Biologicals 32:1727.
14. Hoffmaster, A. R.,, R. F. Meyer,, M. D. Bower,, C. K. Marston,, R. S. Weyant,, K. Thurman,, S. L. Messenger,, E. E. Minor,, J. M. Winchell,, M. V. Rassmussen,, B. R. Newton,, J. T. Parker,, W. E. Morrill,, N. McKinney,, G. A. Barnett,, J. J. Sejvar,, J. A. Jernigan,, B. A. Perkins, and , T. Popovic. 2002. Evaluation and validation of a real-time polymerase chain reaction assay for rapid identification of Bacillus anthracis. Emerg. Infect. Dis. 8:11781182.
15. Hoffmaster, A. R.,, J. Ravel,, D. A. Rasko,, G. D. Chapman,, M. D. Chute,, C. K. Marston,, B. K. De,, C. T. Sacchi,, C. Fitzgerald,, L. W. Mayer,, M. C. Maiden,, F. G. Priest,, M. Barker,, L. Jiang,, R. Z. Cer,, J. Rilstone,, S. N. Peterson,, R. S. Weyant,, D. R. Galloway,, T. D. Read,, T. Popovic, and , C. M. Fraser. 2004. Identification of anthrax toxin genes in a Bacillus cereus associated with an illness resembling inhalation anthrax. Proc. Natl. Acad. Sci. USA. 101:84498454.
16. Jernigan, D. B.,, P. L. Raghunathan,, B. P. Bell,, R. Brechner,, E. A. Bresnitz,, J. C. Butler,, M. Cetron,, M. Cohen,, T. Doyle,, M. Fischer,, C. Greene,, K. S. Griffith,, J. Guarner,, J. L. Hadler,, J. A. Hayslett,, R. Meyer,, L. R. Petersen,, M. Phillips,, R. Pinner,, T. Popovic,, C. P. Quinn,, J. Reefhuis,, D. Reissman,, N. Rosenstein,, A. Schuchat,, W. J. Shieh,, L. Siegal,, D. L. Swerdlow,, F. C. Tenover,, M. Traeger,, J. W. Ward,, I. Weisfuse,, S. Wiersma,, K. Yeskey,, S. Zaki,, D. A. Ashford,, B. A. Perkins,, S. Ostroff,, J. Hughes,, D. Fleming,, J. P. Koplan,, J. L. Gerberding, and The National Anthrax Epidemiologic Investigation Team. 2002. Investigation of bioterrorism-related anthrax, United States, 2001: epidemiologic findings. Emerg. Infect. Dis. 8:10191028.
17. Keim, P.,, A. Kalif,, J. Schupp,, K. Hill,, S. E. Travis,, K. Richmond,, D. M. Adair,, M. Hugh-Jones,, C. R. Kuske, and , P. Jackson. 1997. Molecular evolution and diversity in Bacillus anthracis as detected by amplified fragment length polymorphism markers. J. Bacteriol. 179:818824.
18. Knisely, R. F. 1966. Selective medium for Bacillus anthracis. J. Bacteriol. 92:784786.
19. Little, S. F.,, B. E. Ivins,, P. F. Fellows,, M. L. Pitt,, S. L. Norris, and , G. P. Andrews. 2004. Defining a serological correlate of protection in rabbits for a recombinant anthrax vaccine. Vaccine 22:422430.
20. Logan, N. A., and , P. C. Turnbull. 1999. Bacillus and recently derived genera, p. 357369. In P. R. Murray,, E. J. Baron,, M. A. Pfaller,, F. C. Tenover, and , R. H. Yolken (ed.), Manual of Clinical Microbiology, 7th ed. ASM Press, Washington, D.C.
21. McChlery, S. M., and , S. C. Clarke. 2003. The use of hydrolysis and hairpin probes in real-time PCR. Mol. Biotechnol. 25:26727’4.
22. Morse, S. A.,, R. B. Kellogg,, S. Perry,, R. F. Meyer,, D. Bray,, D. Nichelson, and , J. M. Miller. 2003. Detecting bio-threat agents: the laboratory response network. ASM News 69:433437.
23. Okinaka, R. T.,, K. Cloud,, O. Hampton,, A. R. Hoffmaster,, K. K. Hill,, P. Keim,, T. M. Koehler,, G. Lamke,, S. Kumano,, J. Mahillon,, D. Manter,, Y. Martinez,, D. Ricke,, R. Svensson, and , P. J. Jackson. 1999. Sequence and organization of pXO1, the large Bacillus anthracis plas-mid harboring the anthrax toxin genes. J. Bacteriol. 181:65096515.
24. Peruski, A. H., and , L. F. Peruski, Jr. 2003. Immunological methods for detection and identification of infectious disease and biological warfare agents. Clin. Diagn. Lab. Immunol. 10:506513.
25. Quinn, C. P.,, V. A. Semenova,, C. M. Elie,, S. Romero-Steiner,, C. Greene,, H. Li,, K. Stamey,, E. Steward-Clark,, D. S. Schmidt,, F. Mothershed,, J. Pruckler,, S. Schwartz,, R. F. Benson,, L. O. Helsel,, P. F. Holder,, S. E. Johnson,, M. Kellum,, T. Messmer,, W. L. Thacker,, L. Besser,, B. D. Plikaytis,, T. H. Taylor, Jr.,, A. F. Freeman,, K. J. Wallace,, P. Dull,, J. Sejvar,, F. Bruce,, R. Moreno,, A. Schuchat,, J. R. Lingappa,, S. K. Martin,, J. Walls,, M. Bronsdon,, G. M. Carlone,, M. Bajani-Ari,, D. A. Ashford,, D. S. Stephens, and , B. A. Perkins. 2002. Specific, sensitive, and quantitative enzyme-linked immunosorbent assay for human immunoglobulin G antibodies to anthrax toxin protective antigen. Emerg. Infect. Dis. 8:11031110.
26. Read, T. D.,, S. N. Peterson,, N. Tourasse,, L. W. Baillie,, I. T. Paulsen,, K. F. Nelson,, H. Tettelin,, D. F. Fouts,, J. A. Eisen,, S. R. Gill,, E. K. Holtzapple,, O. A. Okstad,, F. Helgason,, J. Rilstone,, M. Wu,, J. F. Kolonay,, M. J. Beanan,, R. J. Dodson,, L. M. Brinkac,, M. Gwinn,, R. T. DeBoy,, R. Madpu,, S. C. Daugherty,, A. S. Durkin,, D. H. Haft,, W. C. Nelson,, J. D. Peterson,, M. Pop,, H. M. Khouri,, D. Radune,, J. L. Benton,, Y. Mahamoud,, L. Jiang,, I. R. Hance,, J. F. Weidman,, K. J. Berry,, R. D. Plaut,, A. M. Wolf,, K. L. Watkins,, W. C. Nierman,, A. Hazen,, R. Cline,, C. Redmond,, J. F. Thwaite,, O. White,, S. L. Salzberg,, B. Thomason,, A. M. Friedlander,, T. M. Koehler,, P. C. Hanna,, A. B. Kolsto, and , C. M. Fraser. 2003. The genome sequence of Bacillus anthracis Ames and comparison to closely related bacteria. Nature 423:8186.
27. Reif, T. C.,, M. Johns,, S. D. Pillai, and , M. Carl. 1994. Identification of capsule-forming Bacillus anthracis spores with PCR and a novel dual-probe hybridization format. Appl. Environ. Microbiol. 60:16221625.
28. Shieh, W. J.,, J. Guarner,, C. Paddock,, P. Greer,, K. Tatti,, M. Fischer,, M. Layton,, M. Philips,, F. Bresnitz,, C. P. Quinn,, T. Popovic,, B. A. Perkins, and , S. R. Zaki. 2003. The critical role of pathology in the investigation of bioterror-ism-related cutaneous anthrax. Am. J. Pathol. 163:19011910.
29. Ticknor, L. O.,, A. B. Kolsto,, K. K. Hill,, P. Keim,, M. T. Laker,, M. Tonks, and , P. J. Jackson. 2001. Fluorescent amplified fragment length polymorphism analysis of Norwegian Bacillus cereus and Bacillus thuringiensis soil isolates. Appl. Environ. Microbiol. 67:48634873.
30. Turnbull, P. C. 1999. Definitive identification of Bacillus anthracis—a review. J. Appl. Microbiol. 87:237240.


Generic image for table

Diagnostic tests for detection of

Citation: Hoffmaster A, Popovic T. 2006. Detection and Anthrax Diagnosis, p 547-552. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch63

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error