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.
M. avium Mtb Mycobacterium tuberculosis M. avium M. intracellulare M. kansasii M. gordonae M. marinum M. tuberculosis Mycobacterium M. tuberculosis Mtb M. abscessus M. kansasii M. avium M. avium M. intracellulare M. gordonae M. xenopi M. kansasii M. abscessus M. fortuitum M. gordonae M. xenopi Mycobacterium avium Mycobacterium kansasii Mycobacterium abscessus M. abscessus Mtb Mtb Mtb M. kansasii M. marinum secA1 secA1 Mtb Mtb M. bovis Mtb Mtb Mtb M. marium Mycobacterium M. marium M. avium M. intracellulare M. kansasii M. tuberculosis Mtb Mtb Mtb M. abscessus M. avium M. intracellulare M. kansasii M. abscessus M. avium M. PLoS One Eur Respir J J Thorac Dis Ann Am Thorac Soc Int J Infect Dis Tuberc Respir Dis (Seoul) J Clin Microbiol Sci Rep PLoS One Clin Proteom J Clin Microbiol Expert Opin Pharmacother Am J Respir Crit Care Med Infect Immun BMC Evol Biol PLoS Pathog Am J Respir Crit Care Med Bioinformatics Int J Syst Evol Microbiol J Water Health Int J Syst Evol Microbiol Front Cell Infect Microbiol J Clin Microbiol Front Microbiol Int J Dermatol Nat Genet Clin Infect Dis
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