Abstract
In this study, a Liquid Chromatography-Mass Spectrometry (LC-MS) method for the identification of clinically relevant Mycobacteroides abscessus (Mabs) complex organisms is tested using a set of microbial Type strains. This methodology is based on profiling proteins derived from Mycobacteroides abscessus complex isolates. These protein profiles are then used as markers of species differentiation. To test the resolving power, speed, and accuracy of this assay four ATCC type strains and 32 recent clinical isolates of closely related Mabs species collected at ARUP laboratories (10 clinical isolate strains of M. abscessus subsp. abscessus, 10 M. abscessus subsp. massiliense, 2 M. abscessus subsp. bolletii and 10 M. chelonae) were subjected to this approach. Using multiple deconvolution algorithms, we identified hundreds of individual proteins, with subpopulations of these used as species-specific markers. This assay identified 150, 130, 140 and 110 proteoforms with isocratic elution and 230, 180, 200 and 190 proteoforms with gradient elution for M. abscessus (ATCC 19977), M. massiliense (DSM 45103), M. bolletii (DSM 45149) and M. chelonae (ATCC 35752) respectively. Taxonomic species were identified correctly down to the species level with 100% accuracy. The ability to differentiate Mycobacteroides abscessus complex at sub-species level can in-turn be helpful for patient management. Data analysis showed ~7-17 proteoforms potentially able to differentiate between subspecies. Here, we present a proof-of-principle study employing a rapid mass spectrometry-based method to identify the clinically most common species within the Mabs species complex.
Highlights
Growing mycobacteria (RGM) are present everywhere in the environment and cause number of infections in humans such as catheter infection, skin and soft tissue infection, pulmonary infection, disseminated infection, etc
We have previously demonstrated the use of ultra-high-resolution mass spectrometry technique for identification of closely related mycobacterium tuberculosis complex species but no other report yet outlines this technique for Mycobacteroides abscessus (Mabs) complex identification (Bajaj et al, 2021)
Mycobacterium strains belonging to the species M. abscessus, M. massiliense, M. bolletti and M. chelonae were selected as representatives for the Mabs complex
Summary
Growing mycobacteria (RGM) are present everywhere in the environment and cause number of infections in humans such as catheter infection, skin and soft tissue infection, pulmonary infection, disseminated infection, etc. ("Diagnosis and treatment of disease caused by nontuberculous mycobacteria This official statement of the American Thoracic Society was approved by the Board of Directors, March 1997. M. abscessus complex (Mabs complex) comprises several closely related species: M. abscessus, M. massiliense, M. bolletii, M. chelonae, M. franklinii, M. imunogenum, M. salmoniphilum and M. saopaulense responsible for human and zoonotic infections (Adékambi et al, 2004; Adékambi et al, 2006; Bryant et al, 2013). M. bolletti is an uncommon pathogen and is shown to be highly resistant to clarithromycin (Kim et al, 2008; Adékambi and Drancourt, 2009; Koh et al, 2009).In contrast, M. massiliense is susceptible to clarithromycin as it has a non-functional erm(41) gene (Kim et al, 2010). M. chelonae causes infections of human skin and soft tissues and does not contain the erm(41) gene and is susceptible to macrolide treatment. Considering the variations in resistance to the antibiotic therapy and response of a patient to the treatment, it is clinically important to discriminate these closely related species, which will helpful for patient management (Koh et al, 2011; Nakanaga et al, 2011; Harada et al, 2012; Kothavade et al, 2013)
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