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: Kathleen M Tatti, Patricia Greer, Elizabeth White, Wun-Ju Shieh, Jeannette Guarner, Tara Ferebee-Harris, Jeanine Bartlett, David Ashford, Alex Hoffmaster, George Gallucci, Abbas Vafai, Tanja Popovic, Sherif R Zaki
• Journal: Applied immunohistochemistry & molecular morphology : AIMM (Appl Immunohistochem Mol Morphol) Vol. 14 Issue 2 Pg. 234-43 (Jun 2006) ISSN: 1541-2016 [Print] United States
• PMID: 16785797 (Publication Type: Evaluation Study, Journal Article)

Chemical References

• Antibodies, Monoclonal
• Antigens, Bacterial
• Formaldehyde

Topics

• Animals
• Anthrax (diagnosis)
• Antibodies, Monoclonal (analysis)
• Antigens, Bacterial (analysis)
• Bacillus anthracis (cytology, genetics, immunology)
• Formaldehyde (chemistry)
• Humans
• Immunohistochemistry (methods)
• Paraffin Embedding
• Polymerase Chain Reaction
• Staining and Labeling (methods)
• Time Factors
• Tissue Fixation