Electrospray ionization-tandem mass spectrometry analysis of the mycolic acid profiles for the identification of common clinical isolates of mycobacterial species

Rapid method for Mycobacterium tuberculosis identification using electrospray ionization tandem mass spectrometry analysis of mycolic acids

“…Mass spectra analyses presented in this work prove the structure of MAs postulated by other authors. Similar results and mass spectra interpretations based only on precursor ion scans were presented by Song et al, 2009 andShui et al, 2012. The precursor ion scan ( Figure 1A and 1B) showed that the C26 chain is the dominant α-alkyl chain in the mycolic acid profile of Mtb H37Rv.…”

“…The precursor ion scan ( Figure 1A and 1B) showed that the C26 chain is the dominant α-alkyl chain in the mycolic acid profile of Mtb H37Rv. Although there are many recognized species of Mycobacteria the domination of the C26 chain seems to be characteristic of Mtb which cause active TB in humans (Song et al, 2009) therefore it is possible to differentiate between Mtb and MOTT group only the basis of the α-mycolic chain length. In our method MRM pairs including a longer (C26) α-mycolic chain (Q3 mass -395 m/z) were used to confirm the presence of Mtb while MRM pairs built on a shorter (C24) α-mycolic chain determination (Q3 mass -367 m/z) let us check the presence of MOTT strains.…”

“…However, this method is quite sophisticated and expensive relative to other laboratory techniques and is not commonly used in many reference laboratories. Moreover, it requires multiple pretreatment steps for lipid purification and a constant retention time for accurate results (Butler and Guthertz, 2001) More precise methods of mycolic acid analysis involve rapid, high-throughput mass spectrometry techniques that produce both qualitative and quantitative results (Shui et al, 2012;Song et al 2009). In this work, we focused on the development of a rapid ESI-MS/MS technique for the identification and confirmation of TB infection on a triple-quadrupole instrument equipped with an ESI ion source and standard HPLC system.…”

Characterization of Mycolic Acids in Total Fatty Acid Methyl Ester Fractions from <i>Mycobacterium</i> Species by High Resolution MALDI-TOFMS

“…Electrospray ionization (ESI)-tandem mass spectrometry (MS/MS) was applied for the identi cation of common clinical isolate of mycobacterial species. 21) By using tandem mass spectrometry, not only molecular weight pro les but also molecular structural information was provided. is method would be reliable and potentially practical as an identication method for mycobacterial species.…”

Novel phage display-derived mycolic acid-specific antibodies with potential for tuberculosis diagnosis

“…ELISA plates were washed four times in PBS (for polyclonal phage ELISA) or PBS/0.05% Tween-20 (for monoclonal IgG ELISA) and secondary antibody: ␣ M13-HRP against phage major coat protein VIII (GE Healthcare) for phage ELISA or ␣ -human IgG-Fc HRP (Thermo Scientifi c) for monoclonal IgG ELISA, added at 1:5000 dilution in 2% MPBS. Plates were washed as above and one additional time with PBS and then color developed with TMB (Thermo carried out on lipid extracted from such samples using high-performance liquid chromatography, electrospray ionization mass spectrometry, thin layer chromatography, or nuclear magnetic resonance ( 8,(11)(12)(13)(14). However, these methods require highly sophisticated instruments and trained personnel and are not ideal for diagnostic assays in resource-poor settings.…”