title: Morphologic, immunologic, and molecular methods to detect bacillus anthracis in formalin-fixed tissues.
abstract: Due to the importance of Bacillus anthracis as a cause of naturally occurring infection among humans and as an agent of bioterrorism, there is a vital need for rapid and specific assays, including immunohistochemistry (IHC) and polymerase chain reaction (PCR) assays, to detect the bacterium in formalin-fixed tissues. Colorimetric IHC assays were developed using a multistep indirect immunoalkaline phosphatase method with anti-B. anthracis cell wall (EAII-6G6-2-3) and anti-B. anthracis capsule (FDF-1B9) mAbs to detect B. anthracis antigens in formalin-fixed, paraffin-embedded bacterial cultures and tissues. B. anthracis antigens were localized, using both antibodies, in samples from B. anthracis-infected animals and humans. The colorimetric IHC assay with both antibodies was expedient in diagnosing the presence of B. anthracis in formalin-fixed, paraffin-embedded tissue from bioterrorism-associated cases of inhalational and cutaneous anthrax and from a case of naturally occurring cutaneous anthrax. Using the same antibodies, confocal microscopy demonstrated the structure of replicating B. anthracis in tissues. B. anthracis-specific primers were successfully used with PCR to amplify and detect B. anthracis sequences derived from formalin-fixed tissues of anthrax cases. In this study, morphologic, immunologic, and molecular assays were used to study and diagnose 22 veterinary and human anthrax cases.
authors.affiliation: Centers for Disease Control and Prevention, National Center for Infectious Diseases, Division of Viral and Rickettsial Disease, Infectious Disease Pathology Activity, Atlanta, GA, USA.
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Tatti
authors.forename: Patricia
authors.initials: P
authors.lastname: Greer
authors.forename: Elizabeth
authors.initials: E
authors.lastname: White
authors.forename: Wun-Ju
authors.initials: WJ
authors.lastname: Shieh
authors.forename: Jeannette
authors.initials: J
authors.lastname: Guarner
authors.forename: Tara
authors.initials: T
authors.lastname: Ferebee-Harris
authors.forename: Jeanine
authors.initials: J
authors.lastname: Bartlett
authors.forename: David
authors.initials: D
authors.lastname: Ashford
authors.forename: Alex
authors.initials: A
authors.lastname: Hoffmaster
authors.forename: George
authors.initials: G
authors.lastname: Gallucci
authors.forename: Abbas
authors.initials: A
authors.lastname: Vafai
authors.forename: Tanja
authors.initials: T
authors.lastname: Popovic
authors.forename: Sherif R
authors.initials: SR
authors.lastname: Zaki
mesh_terms: D000818:Animals; D000881:Anthrax / Q000175:diagnosis*; D000911:Antibodies, Monoclonal / Q000032:analysis; D000942:Antigens, Bacterial / Q000032:analysis; D001408:Bacillus anthracis / Q000166:cytology* / Q000235:genetics / Q000276:immunology; D005557:Formaldehyde / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D013194:Staining and Labeling / Q000379:methods*; D013997:Time Factors; D016707:Tissue Fixation
chemical_list: D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D005557:Formaldehyde