1 research outputs found
ΠΠ·ΡΡΠ΅Π½ΠΈΠ΅ Π²ΠΈΠ΄ΠΎΠ²ΠΎΠ³ΠΎ ΡΠ°Π·Π½ΠΎΠΎΠ±ΡΠ°Π·ΠΈΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΡΠΎΠ΄Π° Bifidobacterium ΠΊΠΈΡΠ΅ΡΠ½ΠΎΠΉ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΌΠ΅ΡΠΎΠ΄Π° MALDI-TOF ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΠΈ
Background: The members of genus Bifidobacterium represent a significant part of intestinal microbiota in adults and predominate in infants. Species repertoire of the intestinal bifidobacteria is known to be subjected to major changes with age; however, many details of this process are still to be elucidated.Objective: Our aim was to study the diversity of intestinal bifidobacteria and changes of their qualitative and quantitative composition characteristics during the process of growing up using MALDI-TOF mass-spectrometric analysis of pure bacterial cultures.Methods: A cross-sectional study of bifidobacteria in the intestinal microbiota was performed in 93 healthy people of the ages from 1 month to 57 years. Strains were identified using Microflex LT MALDI-TOF MS, the confirmation was performed by 16S rRNA gene fragment sequencing.Results: 93% of isolated bifidobacterial strains were successfully identified using MALDI-TOF mass-spectrometry. At least two of the strains from each species were additionally identified by 16S rRNA gene fragment sequencing, in all of the cases the results were the same. It was shown that the total concentration of bifidobacteria decreases with age (p 0.001) as well as the frequency of isolation of Bifidobacterium bifidum (p =0.020) and Bifidobacterium breve (p 0.001), and the frequency of isolation of Bifidobacterium adolescentis, increases (p 0.001), representing the continuous process of transformation of microbiota.Conclusion: The method of MALDI-TOF mass spectrometry demonstrated the ability to perform rapid and reliable identification of bifidobacteria that allowed the study of changes in the quantitative and qualitative characteristics of human microbiota in the process of growing up.ΠΡΠ΅Π΄ΡΡΠ°Π²ΠΈΡΠ΅Π»ΠΈ ΡΠΎΠ΄Π° Bifidobacterium ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΡΡ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΡΡ ΡΠ°ΡΡΡ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° Π²Π·ΡΠΎΡΠ»ΡΡ
Π»ΡΠ΄Π΅ΠΉ ΠΈ ΡΠΈΡΠ»Π΅Π½Π½ΠΎ Π΄ΠΎΠΌΠΈΠ½ΠΈΡΡΡΡ Π² ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΠ΅ ΠΌΠ»Π°Π΄Π΅Π½ΡΠ΅Π². ΠΠ·Π²Π΅ΡΡΠ½ΠΎ, ΡΡΠΎ Π²ΠΈΠ΄ΠΎΠ²ΠΎΠΉ ΡΠΎΡΡΠ°Π² ΠΊΠΈΡΠ΅ΡΠ½ΡΡ
Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΠΏΠΎΠ΄Π²Π΅ΡΠ³Π°Π΅ΡΡΡ ΡΠΈΠ»ΡΠ½ΡΠΌ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡΠΌ Ρ Π²ΠΎΠ·ΡΠ°ΡΡΠΎΠΌ, ΠΎΠ΄Π½Π°ΠΊΠΎ ΠΌΠ½ΠΎΠ³ΠΈΠ΅ Π΄Π΅ΡΠ°Π»ΠΈ ΡΡΠΎΠ³ΠΎ ΠΏΡΠΎΡΠ΅ΡΡΠ° ΠΎΡΡΠ°ΡΡΡΡ Π½Π΅ΡΡΠ½ΡΠΌΠΈ.Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ: ΠΈΠ·ΡΡΠΈΡΡ Π²ΠΈΠ΄ΠΎΠ²ΠΎΠ΅ ΡΠ°Π·Π½ΠΎΠΎΠ±ΡΠ°Π·ΠΈΠ΅ Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° ΠΈ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΠΈΡ
ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ³ΠΎ ΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ³ΠΎ ΡΠΎΡΡΠ°Π²Π° Π² ΠΏΡΠΎΡΠ΅ΡΡΠ΅ Π²Π·ΡΠΎΡΠ»Π΅Π½ΠΈΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° ΠΏΡΠΈ ΠΏΠΎΠΌΠΎΡΠΈ ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΠΈ MALDI-TOF ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π°Π½Π°Π»ΠΈΠ·Π° Π±Π΅Π»ΠΊΠΎΠ²ΡΡ
ΠΏΡΠΎΡΠΈΠ»Π΅ΠΉ ΡΠΈΡΡΡΡ
ΠΊΡΠ»ΡΡΡΡ.ΠΠ΅ΡΠΎΠ΄Ρ: ΠΊΡΠΎΡΡ-ΡΠ΅ΠΊΡΠΈΠΎΠ½Π½ΠΎΠ΅ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ ΡΠ°Π·Π½ΠΎΠΎΠ±ΡΠ°Π·ΠΈΡ Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ Π² ΡΠΎΡΡΠ°Π²Π΅ Π½ΠΎΡΠΌΠ°Π»ΡΠ½ΠΎΠΉ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ΠΎ Ρ 93 ΡΠ΅Π»ΠΎΠ²Π΅ΠΊ Π² Π²ΠΎΠ·ΡΠ°ΡΡΠ΅ ΠΎΡ 1 ΠΌΠ΅Ρ Π΄ΠΎ 57 Π»Π΅Ρ. ΠΡΡΡΠ΅ΡΡΠ²Π»ΡΠ»ΠΈ Π²ΡΠ΄Π΅Π»Π΅- Π½ΠΈΠ΅ ΡΠΈΡΡΡΡ
ΠΊΡΠ»ΡΡΡΡ ΠΈ ΠΈΡ
ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΡ Π½Π° ΠΏΡΠΈΠ±ΠΎΡΠ΅ Microflex LT MALDI-TOF MS (Bruker Daltonics, ΠΠ΅ΡΠΌΠ°Π½ΠΈΡ), ΠΏΠΎΠ΄ΡΠ²Π΅ΡΠΆΠ΄Π΅Π½ΠΈΠ΅ ΡΠ΅Π°Π»ΠΈΠ·ΠΎΠ²ΡΠ²Π°Π»ΠΈ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠ° Π³Π΅Π½Π° 16S ΡΠ ΠΠ.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: Ρ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ΠΌ MALDI-TOF ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΠΈ Π±ΡΠ»ΠΎ ΡΡΠΏΠ΅ΡΠ½ΠΎ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½ΠΎ 93% Π²ΡΠ΄Π΅Π»Π΅Π½Π½ΡΡ
ΡΡΠ°ΠΌΠΌΠΎΠ² Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ. ΠΠΈΠ½ΠΈΠΌΡΠΌ ΠΏΠΎ 2 ΠΏΡΠ΅Π΄ΡΡΠ°Π²ΠΈΡΠ΅Π»Ρ ΠΎΡ ΠΊΠ°ΠΆΠ΄ΠΎΠ³ΠΎ ΠΈΠ· Π²ΠΈΠ΄ΠΎΠ² Π±ΡΠ»ΠΈ Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΠΎ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½Ρ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠ° Π³Π΅Π½Π° 16SΡΠ ΠΠ; Π²ΠΎ Π²ΡΠ΅Ρ
ΡΠ»ΡΡΠ°ΡΡ
ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΡ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ² ΡΠΎΠ²ΠΏΠ°Π»ΠΈ. ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ Ρ Π²ΠΎΠ·ΡΠ°ΡΡΠΎΠΌ ΠΏΡΠΎΠΈΡΡ
ΠΎΠ΄ΠΈΡ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΠ΅ ΠΎΠ±ΡΠ΅ΠΉ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ (p 0,001), ΡΠΌΠ΅Π½ΡΡΠ°Π΅ΡΡΡ Π²ΡΡΡΠ΅ΡΠ°Π΅ΠΌΠΎΡΡΡ Π²ΠΈΠ΄ΠΎΠ² Bifidobacterium bifidum (p =0,020) ΠΈ Bifidobacterium breve (p 0,001), Π° Π²ΡΡΡΠ΅ΡΠ°Π΅ΠΌΠΎΡΡΡ Π²ΠΈΠ΄Π° Bifidobacterium adolescentis ΡΠ²Π΅Π»ΠΈΡΠΈΠ²Π°Π΅ΡΡΡ (p 0,001), ΠΎΡΡΠ°ΠΆΠ°Ρ ΠΏΠΎΡΡΠ΅ΠΏΠ΅Π½Π½ΡΠ΅ ΠΏΡΠΎΡΠ΅ΡΡΡ ΠΏΠ΅ΡΠ΅ΡΡΡΠΎΠΉΠΊΠΈ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ.ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅: ΠΌΠ΅ΡΠΎΠ΄ MALDI-TOF ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΠΈ ΠΏΠΎΠΊΠ°Π·Π°Π» Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΡ Π±ΡΡΡΡΠΎΠΉ ΠΈ Π½Π°Π΄Π΅ΠΆΠ½ΠΎΠΉ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ, ΠΏΠΎΠ·Π²ΠΎΠ»ΠΈΠ²ΡΠ΅ΠΉ ΠΏΡΠΎΠ²Π΅ΡΡΠΈ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠΉ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΈ ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Π΅ΠΉ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° Π² ΠΏΡΠΎΡΠ΅ΡΡΠ΅ Π²Π·ΡΠΎΡΠ»Π΅Π½ΠΈ