Characterization and Differentiation of Mycobacterium avium subsp. paratuberculosis from Other Mycobacteria Using Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Abstract

Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne's disease in cattle, is responsible for significant economic losses to the US dairy industry. The pathogen has also been associated with chronic human diseases like Crohn's disease, type 1 diabetes and multiple sclerosis. Determining causation requires rapid characterization and source tracking the pathogen. Here, we used matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to characterize and differentiate strains of MAP from 14 other species of Mycobacterium from bovine, human, and environmental sources. Lysates from cells disrupted by bead beating in TFA-acetonitrile solution were analyzed by MALDI-TOF. MAP strains were differentiated by mass spectral profiles that are distinct from each other and from other Mycobacterium species. Cluster analysis of spectral profiles indicates two distinct clusters, one dominated by the members of avium complex and a second group dominated by members of fortuitum and parafortuitum complexes. We believe that MALDI-TOF methods can be used to differentiate and source-track MAP strains.

Mycobacterial Infections in Wild Boars (Sus scrofa) from Southern Switzerland: Diagnostic Improvements, Epidemiological Situation and Zoonotic Potential

“…In most cases, however, the source of infection remained unclear. The discrimination ofMap with the MALDI-TOF technique has been assessed in previous studies (Ravva et al, 2017, Ricchi et al, 2017. The 25 MAC isolates tested in the present study were all confirmed to be Mah by sequence analysis, and the first subspecies suggestion provided by MALDI-TOF was generally correct (80%).…”

Mycobacterial infections in wild boars ( Sus scrofa ) from Southern Switzerland: Diagnostic improvements, epidemiological situation and zoonotic potential

“…Although the threshold recommended by the manufacturer enabled the correct identification of only 72.1% of the isolates, a lower cut-off for mycobacteria is commonly used (Alcolea-Medina et al, 2019;Buchan, Riebe, Timke, Kostrzewa, & Ledeboer, 2014;Mediavilla-Gradolph et al, 2015;Saleeb et al, 2011). In the authors' opinion, an important drawback of the MALDI-TOF MS technology in comparison with sequence analysis of housekeeping genes such as 16S rRNA, is that yet undescribed species will go undetected, either without any identification or as misidentifi- (Ravva, Harden, & Sarreal, 2017;Ricchi et al, 2017). The 25 M. avium isolates tested in the present study were all confirmed to be Mah by sequence analysis, and the first subspecies suggestion provided by MALDI-TOF MS was generally correct (80%).…”

Instrumental analytical tools for mycobacteria characterisation

“…Another presented option was a mycobacterial suspension in the liquid culture medium. Frequently used solid culture media were Löwenstein-Jensen medium, Middlebrook 7H10, and Review https://doi.org/10.17221/69/2021-CJFS (Fangous et al 2014) human (bronchoalveolar and lavage, peritoneal fluid, pus, sputum, urine) (Genc et al 2018) laboratory culture collection (Hettick et al 2004) human; laboratory culture collection (Lin et al 2015) laboratory culture collection (Mather et al 2014) human (Machen et al 2013) human (respiratory specimens) (Mediavilla-Gradolph et al 2015) laboratory culture collection (Moreno et al 2018) laboratory culture collection (Neumann et al 2019) human (Park et al 2016) human (sputum, respiratory aspirates); heater-cooler units; laboratory culture collection (Pranada et al 2017) aquarium fish (Puk et al 2018) bovine, chicken, human, swine, wood pigeon; dairy water trough, soil (Ravva et al 2017) human (Rodriguez-Temporal et al 2018) human (Rotcheewaphan et al 2019) human (Tseng et al 2013) Whole cells isolates laboratory culture collection (Hettick et al 2004) human (Larrouy-Maumus and Puzo 2015) tissue sections mouse and rabbit lungs (Blanc et al 2018) https://doi.org/10.17221/69/2021-CJFS 7H11 agar, while the most common liquid culture media were Middlebrook 7H9 broth, Sauton's medium or liquid BD BACTEC mycobacteria growth indicator tubes. The bone powder obtained from archaeological human skeletal remains proved to be also suitable for mycobacteria detection in a few cases.…”