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Alternative protocol leading to rapid identification of Actinomycetes isolated from Algerian desertic soil by MALDI-TOF mass spectrometry.
- Source :
-
Journal of microbiological methods [J Microbiol Methods] 2024 Aug; Vol. 223, pp. 106984. Date of Electronic Publication: 2024 Jun 30. - Publication Year :
- 2024
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Abstract
- Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is the first-line method for the rapid identification of most cultured microorganisms. As for Streptomyces strains, MALDI-TOF MS identification is complicated by the characteristic incrustation of colonies in agar and the strong cell wall of Actinomycetes cells requiring the use of alternative protein extraction protocols. In this study, we developed a specific protocol to overcome these difficulties for the MALDI-TOF MS identification of Actinomycetes made on solid medium. This protocol includes incubation of colony removed from agar plate with the beta-agarase enzyme, followed by a mechanical lysis and two washes by phosphate buffer and ethanol. Twenty-four Streptomyces and two Lentzea strains isolated from Algerian desertic soils were first identified by 16S rRNA sequencing as gold standard method, rpoB gene was used as a secondary gene target when 16S rRNA did not allow species identification. In parallel the isolates were identified by using the MALDI-TOF MS protocol as reported. After the expansion of the database with the inclusion of this MSP <subscript>S</subscript> , the strains were analyzed again in MALDI Biotyper, and all were identified. This work demonstrates that the rapid identification of Actinomycetes can be obtained without protein extraction step frequently used in MALDI-TOF mass spectrometry with this type of microorganisms.<br />Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.<br /> (Copyright © 2024 Elsevier B.V. All rights reserved.)
- Subjects :
- Algeria
DNA, Bacterial genetics
Streptomyces isolation & purification
Streptomyces genetics
Streptomyces classification
Streptomyces chemistry
Bacterial Proteins genetics
DNA-Directed RNA Polymerases genetics
Culture Media chemistry
Sequence Analysis, DNA
Bacteriological Techniques methods
Glycoside Hydrolases
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods
Soil Microbiology
RNA, Ribosomal, 16S genetics
Actinobacteria isolation & purification
Actinobacteria genetics
Actinobacteria classification
Actinobacteria chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1872-8359
- Volume :
- 223
- Database :
- MEDLINE
- Journal :
- Journal of microbiological methods
- Publication Type :
- Academic Journal
- Accession number :
- 38955305
- Full Text :
- https://doi.org/10.1016/j.mimet.2024.106984