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Conjunctival Commensal Isolation and Identification in Mice.

Authors
  • Smith-Page, Kirsten1
  • Kugadas, Abirami1
  • Lin, Tiffany1
  • Delaney, Mary2
  • Bry, Lynn2
  • Gadjeva, Mihaela3
  • 1 Department of Medicine, Division of Infectious Diseases, Brigham and Women's Hospital, Harvard Medical School.
  • 2 Massachusetts Host-Microbiome Center, Department of Pathology, Brigham and Women's Hospital, Harvard Medical School; Clinical Microbiology Laboratory, Department of Pathology, Brigham and Women's Hospital, Harvard Medical School.
  • 3 Department of Medicine, Division of Infectious Diseases, Brigham and Women's Hospital, Harvard Medical School; [email protected]
Type
Published Article
Journal
Journal of Visualized Experiments
Publisher
MyJoVE Corporation
Publication Date
May 01, 2021
Issue
171
Identifiers
DOI: 10.3791/61672
PMID: 33999020
Source
Medline
Language
English
License
Unknown

Abstract

The ocular surface was once considered immune privileged and abiotic, but recently it appears that there is a small, but persistent commensal presence. Identification and monitoring of bacterial species at the ocular mucosa have been challenging due to their low abundance and limited availability of appropriate methodology for commensal growth and identification. There are two standard approaches: culture based or DNA sequencing methods. The first method is problematic due to the limited recoverable bacteria and the second approach identifies both live and dead bacteria leading to an aberrant representation of the ocular space. We developed a robust and sensitive method for bacterial isolation by building upon standard microbiological culturing techniques. This is a swab-based technique, utilizing an "in-lab" made thin swab that targets the lower conjunctiva, followed by an amplification step for aerobic and facultative anaerobic genera. This protocol has allowed us to isolate and identify conjunctival species such as Corynebacterium spp., Coagulase Negative Staphylococcus spp., Streptococcus spp., etc. The approach is suitable to define commensal diversity in mice under different disease conditions.

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