ACIDIC METHANOLYSIS V. ALKALINE SAPONIFICATION IN GAS CHROMATOGRAPHIC CHARACTERIZATION OF MYCOBACTERIA: DIFFERENTIATION BETWEEN <i>MYCOBACTERIUM AVIUM‐INTRACELLULARE</i> AND <i>MYCOBACTERIUM GASTRI</i>

Larsson, Lennart

doi:10.1111/j.1699-0463.1983.tb00039.x

Cited by 6 publications

(1 citation statement)

References 7 publications

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“…Alternatively, the mycobacteria may be subjected to acid methanolysis instead of saponification, because in the former method the methyl esters are formed without any production of free acids. This phenomenon is discussed in more detail elsewhere (8,9).…”

Section: Discussionmentioning

confidence: 81%

Influence of Culture Medium and Incubation Time on the Fatty Acid Compositions of Some Rapid‐Growing Mycobacteria as Analysed by Packed and Capillary Column gas Chromatography

Larsson

Valero-Guillén

Martin-Luengo

et al. 1985

Acta Pathologica Microbiologica Scandinavica Series B: Microbio

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Cellular fatty acids of four rapid‐growing mycobacterial species (Mycobacterium chelonei, M. fortuitum, M. phlei, and M. smegmatis) were analysed by packed and capillary column gas chromatography after one, three, four, six, eight, and twelve days of incubation on Löwenstein‐Jensen and Sauton agar media. Variations in incubation time did not affect the chromatograms except in the case of twelve‐day incubated M. smegmatis. Mycobacteria cultivated on Sauton agar medium contained more tuberculostearic and less oleic acid compared with Löwenstein‐Jensen. For informative and reproducible analytical results, we recommend using a chemically defined culture medium and splitless injection on a capillary column capable of separating not only the methyl esters of the cellular fatty acids but also the diagnostically important secondary alcohols containing 18 and 20 carbon atoms.

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Section: Discussionmentioning

confidence: 81%

Influence of Culture Medium and Incubation Time on the Fatty Acid Compositions of Some Rapid‐Growing Mycobacteria as Analysed by Packed and Capillary Column gas Chromatography

Larsson

Valero-Guillén

Martin-Luengo

et al. 1985

Acta Pathologica Microbiologica Scandinavica Series B: Microbio

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Gas chromatographic fatty acid profiles for characterisation of mycobacteria: An interlaboratory methodological evaluation

Larsson

Jantzen²,

Johnsson

1985

Eur. J, Clin. Microbiol.

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Three species of mycobacteria were cultured and processed for cellular fatty acid analysis by capillary gas chromatography in three laboratories to study interlaboratory variations of the resulting chromatographic profiles. Largely consistent and characteristic fatty acid profiles were obtained, although there were minor quantitative variations in the patterns due to methodological differences (cultivation, hydrolysis, derivatization, gas chromatographic conditions etc.). The following points were important for achieving informative and reproducible results. A chemically defined growth medium (e.g., Proskauer-Beck) provides more consistent profiles than the lipid-rich Löwenstein-Jensen medium. Harvesting directly into the digesting solution (NaOH or HCl in methanol) followed by heating or autoclaving is a simple and reliable way of releasing fatty acids. Care should be taken to ensure reproducible detection of long-chain alcohols either by using acid methanolysis or including a base-wash step in the procedure following alkaline hydrolysis. The temperature of the gas chromatographic injector should be at least 325 degrees C. A capillary column of a minimum length of 10 m coated with a methyl silicone is adequate. Our results indicate the possibility of recommending a practical and reproducible gas chromatographic procedure for mycobacterial characterisation.

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Faster identification of mycobacteria using gas liquid and thin layer chromatography

Parez

Fauville-Dufaux²,

Dossogne

et al. 1994

Eur. J. Clin. Microbiol. Infect. Dis.

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Gas liquid chromatography (GLC) and thin layer chromatography (TLC) analysis of cell wall content was used for identification of mycobacteria isolated in primary cultures. GLC permitted determination of the fatty acid and alcohol profiles of Mycobacterium simiae and Mycobacterium marinum and detection of a peak in Mycobacterium ulcerans formerly described for Mycobacterium malmoense. Using the data obtained to fill some of the gaps in the dichotomic trees of Tisdall et al. and Jantzen et al., GLC analysis allowed full identification of 8 of 22 mycobacterial species after 24 hours. The other 14 species could be divided into four groups on the basis of similar findings on GLC. TLC was used for full identification of three species. The identification results of conventional methods were concordant with those of GLC and TLC in 161 of 169 strains (93%) representing 21 different species. Using primarily chromatography for analysis of cell wall content, and in the case of some species complementary biochemical tests, the identification procedure could be shortened to a maximum of three days after primary culture.

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ACIDIC METHANOLYSIS V. ALKALINE SAPONIFICATION IN GAS CHROMATOGRAPHIC CHARACTERIZATION OF MYCOBACTERIA: DIFFERENTIATION BETWEEN MYCOBACTERIUM AVIUM‐INTRACELLULARE AND MYCOBACTERIUM GASTRI

Cited by 6 publications

References 7 publications

Influence of Culture Medium and Incubation Time on the Fatty Acid Compositions of Some Rapid‐Growing Mycobacteria as Analysed by Packed and Capillary Column gas Chromatography

Influence of Culture Medium and Incubation Time on the Fatty Acid Compositions of Some Rapid‐Growing Mycobacteria as Analysed by Packed and Capillary Column gas Chromatography

Gas chromatographic fatty acid profiles for characterisation of mycobacteria: An interlaboratory methodological evaluation

Faster identification of mycobacteria using gas liquid and thin layer chromatography

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