Peptidomic analysis of mycobacterial secreted proteins enables species identification
Abstract
Pulmonary disease arising from slow‐growing mycobacterial infections has emerged as an increasingly prevalent clinical concern over the past two to three decades. Proteins belonging to the family of ESAT‐6 secretion (Esx) systems play critical roles in the virulence of most pathogenic mycobacterial species and are associated with drug resistance. However, no clinical applications can detect and discriminate the expression of species‐specific variants of these proteins in clinical samples, such as early growth cultures, for rapid diagnosis of specific mycobacterial infections, which may require distinct interventions. Conventional immunoassay approaches are not suitable for this purpose due to the significant degree of conservation of Esx proteins among species. Herein we describe the development of a novel immunoprecipitation‐coupled mass spectrometry assay that can distinguish Esx proteins that are expressed by slow‐growing mycobacterial species commonly detected in clinical isolates. This approach uses custom antibodies raised against single semi‐conserved peptide regions in M. tuberculosis (Mtb) EsxB and EsxN to capture corresponding peptides from protein orthologs of mycobacteria associated with human respiratory infections, including Mtb, M. avium, M. intracellulare, M. kansasii, M. gordonae, and M. marinum, to detect these species in standard clinical cultures at the first sign mycobacterial growth to allow rapid disease diagnosis.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Apr 18, 2022
- Source ID
- 10.1002/viw.20210019
Entities
People
- Adrian M. Zelazny
- Christopher J. Lyon
- Jia Fan
- Lingpeng Zhan
- Meena Rajagopal
- Qingbo Shu
- Suwatchareeporn Rotcheewaphan
- Tony Hu
- Wei Sha
- Xiangxing Kong
- Yifan He
Organizations
- Chulalongkorn University
- National Institutes of Health
- National Institutes of Health Clinical Center
- Tongji University
- Tulane University of Louisiana
- United States Department of Defense