Evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid-resistant bacteria of the order actinomycetales by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Background: Problem with mass spectrometers is the difficulty in identifying some genera of acid-fast bacteria (AFB). Due to the peculiarities of the colony architectonics (the formation of dry colonies with a complex structure) and the structure of the cell wall, the probability of obtaining a suf...

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Main Authors: Artem Victorovich Lyamin, Alena Anatolyevna Ereshchenko, Danir Damirovich Ismatullin, Maxim Olegovich Zolotov, Dmitriy Vladimirovich Alekseev, Karim Askerovich Kayumov
Format: Article
Language:English
Published: Wolters Kluwer Medknow Publications 2023-01-01
Series:International Journal of Mycobacteriology
Subjects:
Online Access:http://www.ijmyco.org/article.asp?issn=2212-5531;year=2023;volume=12;issue=2;spage=157;epage=161;aulast=Lyamin
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author Artem Victorovich Lyamin
Alena Anatolyevna Ereshchenko
Danir Damirovich Ismatullin
Maxim Olegovich Zolotov
Dmitriy Vladimirovich Alekseev
Karim Askerovich Kayumov
author_facet Artem Victorovich Lyamin
Alena Anatolyevna Ereshchenko
Danir Damirovich Ismatullin
Maxim Olegovich Zolotov
Dmitriy Vladimirovich Alekseev
Karim Askerovich Kayumov
author_sort Artem Victorovich Lyamin
collection DOAJ
description Background: Problem with mass spectrometers is the difficulty in identifying some genera of acid-fast bacteria (AFB). Due to the peculiarities of the colony architectonics (the formation of dry colonies with a complex structure) and the structure of the cell wall, the probability of obtaining a sufficient amount of ribosomal proteins is significantly reduced. This problem is partially solved using an extended method of direct application and extraction with formic acid, which can significantly improve the quality of the identification. Methods: The study analyzed strains of microorganisms obtained during the examination of patients with suspected tuberculosis. In total, 287 strains of nontuberculous mycobacteria (NTM) were obtained. In addition, 63 strains of the most common bacteria from the AFB group were analyzed. Matrix-assisted laser desorption/ionization (MALDI) was used. Three main methods of sample preparation of microorganisms according to the manufacturer's recommendations for use with the MALDI-time-of-flight (ToF) mass spectrometry method were used in the work: Direct coating method, extended direct coating method, and formic acid extraction method. Results: When evaluating the influence of the cultivation medium on the result of NTM identification by MALDI-ToF mass spectrometry, statistically significant results of the influence of the medium on the result of NTM identification were revealed for all compared parameters. Conclusions: Optimization of sample preparation protocols and assessment of the impact on the identification of new methods of cultivating microorganisms can significantly improve the quality of the identification of both clinically significant microorganisms from the AFB group and saprophytic microflora, the clinical significance of which has not been proven at the moment.
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spelling doaj.art-9f4926f3d4444cae9db1cfc2cd3246962023-07-23T11:14:42ZengWolters Kluwer Medknow PublicationsInternational Journal of Mycobacteriology2212-55312212-554X2023-01-0112215716110.4103/ijmy.ijmy_85_23Evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid-resistant bacteria of the order actinomycetales by matrix-assisted laser desorption/ionization time-of-flight mass spectrometryArtem Victorovich LyaminAlena Anatolyevna EreshchenkoDanir Damirovich IsmatullinMaxim Olegovich ZolotovDmitriy Vladimirovich AlekseevKarim Askerovich KayumovBackground: Problem with mass spectrometers is the difficulty in identifying some genera of acid-fast bacteria (AFB). Due to the peculiarities of the colony architectonics (the formation of dry colonies with a complex structure) and the structure of the cell wall, the probability of obtaining a sufficient amount of ribosomal proteins is significantly reduced. This problem is partially solved using an extended method of direct application and extraction with formic acid, which can significantly improve the quality of the identification. Methods: The study analyzed strains of microorganisms obtained during the examination of patients with suspected tuberculosis. In total, 287 strains of nontuberculous mycobacteria (NTM) were obtained. In addition, 63 strains of the most common bacteria from the AFB group were analyzed. Matrix-assisted laser desorption/ionization (MALDI) was used. Three main methods of sample preparation of microorganisms according to the manufacturer's recommendations for use with the MALDI-time-of-flight (ToF) mass spectrometry method were used in the work: Direct coating method, extended direct coating method, and formic acid extraction method. Results: When evaluating the influence of the cultivation medium on the result of NTM identification by MALDI-ToF mass spectrometry, statistically significant results of the influence of the medium on the result of NTM identification were revealed for all compared parameters. Conclusions: Optimization of sample preparation protocols and assessment of the impact on the identification of new methods of cultivating microorganisms can significantly improve the quality of the identification of both clinically significant microorganisms from the AFB group and saprophytic microflora, the clinical significance of which has not been proven at the moment.http://www.ijmyco.org/article.asp?issn=2212-5531;year=2023;volume=12;issue=2;spage=157;epage=161;aulast=Lyaminacid-resistant bacteriamatrix-assisted laser desorption/ionization time-of-flight mass spectrometrynontuberculous mycobacteria
spellingShingle Artem Victorovich Lyamin
Alena Anatolyevna Ereshchenko
Danir Damirovich Ismatullin
Maxim Olegovich Zolotov
Dmitriy Vladimirovich Alekseev
Karim Askerovich Kayumov
Evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid-resistant bacteria of the order actinomycetales by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
International Journal of Mycobacteriology
acid-resistant bacteria
matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
nontuberculous mycobacteria
title Evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid-resistant bacteria of the order actinomycetales by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_full Evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid-resistant bacteria of the order actinomycetales by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_fullStr Evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid-resistant bacteria of the order actinomycetales by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_full_unstemmed Evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid-resistant bacteria of the order actinomycetales by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_short Evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid-resistant bacteria of the order actinomycetales by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_sort evaluation of the influence of the cultivation medium on the result of identification of microorganisms from the group of acid resistant bacteria of the order actinomycetales by matrix assisted laser desorption ionization time of flight mass spectrometry
topic acid-resistant bacteria
matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
nontuberculous mycobacteria
url http://www.ijmyco.org/article.asp?issn=2212-5531;year=2023;volume=12;issue=2;spage=157;epage=161;aulast=Lyamin
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