Mycolic Acid Analysis by High-Performance Liquid Chromatography for Identification ofMycobacteriumSpecies

Abstract

Mycobacterium tuberculosis is the etiologic agent of tuberculosis and can be accurately detected by laboratories using commercial genetic tests. Nontuberculosis mycobacteria (NTM) causing other mycobacterioses can be difficult to identify. The identification processes are confounded by an increasing diversity of newly characterized NTM species. The ubiquitous nature of NTM, combined with their potential to be opportunistic pathogens in immunocompromised as well as nonimmunodeficient patients, further complicates the problem of their identification. Since clinical case management varies depending on the etiologic agent, laboratories must identify the species in a timely manner. However, only a few identification methods can detect the species diversity within the Mycobacterium genus. Over the last decade, high-performance liquid chromatography analysis of the mycolic acids has become an accepted method for identification of mycobacteria. In this review, we assess its development and usefulness as an identification technique for Mycobacterium species

Towards understanding the functional diversity of cell wall mycolic acids of Mycobacterium tuberculosis

“…A homologous series of α-mycolates from Mtb H37Ra was purified by HPLC under reverse phase conditions and each component characterised by MS, leading to the identification of 10 components with a range of chain lengths [42]. This has been extended to establish characteristic HPLC patterns that can be used to distinguish a range of mycobacteria [43,44]. For example, the MA of M.bovis BCG can be distinguished from those of M.tuberculosis and M. bovis [45].…”

Mycobacterium shottsii sp. nov., a slowly growing species isolated from Chesapeake Bay striped bass (Morone saxatilis)

“…Specimens were processed as described previously using the standard method for sample preparation and UV analysis (Butler et al, 1996). Differentiation of specific Mycobacterium species was by a visual decision method using an internal size standard, an identification process reviewed recently by Butler & Guthertz (2001).…”

Mycobacterium psychrotolerans sp. nov., isolated from pond water near a uranium mine

“…2). M. sphagni is also found in this group (Butler & Guthertz, 2001). WA101 T can easily be separated from M. sphagni and other species of this group on the basis of quantitative differences.…”

“…WA101 T can easily be separated from M. sphagni and other species of this group on the basis of quantitative differences. Isolate WA101 T was also distinguished from Mycobacterium chubuense, Mycobacterium obuense and M. parafortuitum on the basis of HPLC mycolic acid chromatograms as the latter species present triplepeak clusters, with those in the early cluster emerging before 5 min (Butler & Guthertz, 2001). WA101 T grew between 4 and 37 u C on GYEA; no growth was recorded at 45 or 52 uC.…”