Mycobacterial Cultivation and Strain Preparation

M. abscessus, M. bolletii, and M. massiliense clinical isolates were a kind gift from the National Mycobacterium Reference Laboratory, Borstel, Germany. An auxotrophic mutant strain of M. tuberculosis (H37Rv ΔpanCD ΔleuCD) was used as a control (Vilchèze et al., 2018 (link)). Mycobacterial strains were grown on Middlebrook 7H11 agar (Sigma-Aldrich) supplemented with 0.5% glycerol and 10% oleic albumin dextrose catalase (OADC) (Becton Dickinson (BD), Oxford, UK), and incubated at 37°C for 3 to 5 days for MABC strains and, 14 to 21 days for M. tuberculosis. E. coli BW25141 was grown on LB agar (Sigma-Aldrich) and used as a further control.

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Akwani W.C., van Vliet A.H., Joel J.O., Andres S., Diricks M., Maurer F.P., Chambers M.A, & Hingley-Wilson S.M. (2022). The Use of Comparative Genomic Analysis for the Development of Subspecies-Specific PCR Assays for Mycobacterium abscessus. Frontiers in Cellular and Infection Microbiology, 12, 816615.

Publication 2022

Middlebrook 7H11 agar oleic albumin dextrose catalase (OADC) LB agar

Agar Albumin Catalase Dextrose E coli Glycerol M tuberculosis Mycobacterial Strains

Corresponding Organization : University of Surrey

Other organizations : Research Center Borstel - Leibniz Lung Center, German Center for Infection Research, University Medical Center Hamburg-Eppendorf, Universität Hamburg

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Variable analysis

independent variables

Mycobacterial strains

dependent variables

Growth of mycobacterial strains

control variables

Growth conditions (Middlebrook 7H11 agar, 0.5% glycerol, 10% OADC, 37°C incubation)
Incubation time (3-5 days for MABC strains, 14-21 days for M. tuberculosis)

controls

Positive control: Auxotrophic mutant strain of M. tuberculosis (H37Rv ΔpanCD ΔleuCD)
Negative control: E. coli BW25141

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