Laser Scanning Confocal Microscopy Was Used to Validate the Presence of Burkholderia Pseudomallei or B. Mallei in Formalin-Fixed Paraffin Embedded Tissues

Tropical Medicine and Infectious Disease

WilsonJ. Ribot¹  Kei Amemiya¹  SylviaR. Trevino¹  PatriciaL. Worsham¹  DavidM. Waag¹  SusanL. Welkos¹  ChristopherK. Cote¹  JenniferL. Dankmeyer¹  Carl Soffler¹  JeremyJ. Bearss²  Xiankun Zeng²  RobertC. Bernhards³ 

[1] Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702, USA;Pathology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702, USA;U.S. Army Combat Capabilities Development Command Chemical Biological Center, 8198 Blackhawk Road, Aberdeen Proving Ground, MD 21010, USA;

关键词: Burkholderia pseudomallei;    melioidosis;    Burkholderia mallei;    glanders;    laser scanning confocal microscopy;    formalin-fixed paraffin embedded tissue;   

DOI  :  10.3390/tropicalmed5020065

来源: DOAJ

【 摘 要 】

Burkholderia pseudomallei and B. mallei are Gram-negative, facultative intracellular bacteria that cause melioidosis and glanders, respectively. Currently, there are no vaccines for these two diseases. Animal models have been developed to evaluate vaccines and therapeutics. Tissues from infected animals, however, must be fixed in formalin and embedded in paraffin (FFPE) before analysis. A brownish staining material in infected tissues that represents the exopolysaccharide of the pathogen was seen by bright field microscopy but not the actual microorganism. Because of these results, FFPE tissue was examined by laser scanning confocal microscopy (LSCM) in an attempt to see the microorganism. Archival FFPE tissues were examined from ten mice, and five nonhuman primates after exposure to B. pseudomallei or B. mallei by LSCM. Additionally, a historical spleen biopsy from a human suspected of exposure to B. mallei was examined. B. pseudomallei was seen in many of the infected tissues from mice. Four out of five nonhuman primates were positive for the pathogen. In the human sample, B. mallei was seen in pyogranulomas in the spleen biopsy. Thus, the presence of the pathogen was validated by LSCM in murine, nonhuman primate, and human FFPE tissues.

【 授权许可】

Unknown