Method for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria

The study's purpose was the developing of the method for Acinetobacter nosocomialis bacteria identification by the totality of phenotypic characteristics of the Acinetobacter baumannii (Ab) group based on the urease activity trait.The research's objects were clinical strains of the Ab grou...

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Autores principales:	E. P. Sivolodskii, E. V. Zueva
Formato:	article
Lenguaje:	RU
Publicado:	Sankt-Peterburg : NIIÈM imeni Pastera 2021
Materias:	acinetobacter nosocomialis acinetobacter baumannii bacterial urease rapid activity urease glucose utilization Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/01488526aba4406788a953b529e808d6
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spelling	oai:doaj.org-article:01488526aba4406788a953b529e808d62021-11-22T07:09:54ZMethod for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria2220-76192313-739810.15789/2220-7619-MFP-1422https://doaj.org/article/01488526aba4406788a953b529e808d62021-06-01T00:00:00Zhttps://www.iimmun.ru/iimm/article/view/1422https://doaj.org/toc/2220-7619https://doaj.org/toc/2313-7398The study's purpose was the developing of the method for Acinetobacter nosocomialis bacteria identification by the totality of phenotypic characteristics of the Acinetobacter baumannii (Ab) group based on the urease activity trait.The research's objects were clinical strains of the Ab group, isolated in 2018—2019 in the bacteriological laboratory of Kirov Military Medical Academy (St. Petersburg), of which were A. baumannii (n = 85), A. nosocomialis (n = 12), A. pittii (n = 10). In addition, 9 strains of A. nosocomialis were isolated from water samples from the Neva River and 2 strains of A. calcoaceticus from the cyanobacterial mats. The belonging of the strains to Ab group was determined by the combination of the metabolic and physiological characteristics of the taxonomic tests for this group. The species identification of the studied strains was determined by the matrix-associated laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). The urease activity of bacteria was determined using the microvolume method. We used a medium with urea in the following composition (g/l): Na2HPO4 — 1.1; KH2PO4 — 1.1; NaCI — 5.0; 40% alkaline solution phenol red — 5 ml; urea — 10—15; distilled water — 1 l. The ingredients, without urea, were dissolved in distilled water, dispensed to vials, sterilized for 20 minutes at 121°C, urea was added in the calculated amount. The medium with urea was added in 0.1 ml to wells of the plate, then inoculated the daily agar culture of the studied and control strains by full loop (2 mm in diameter) and incubated under aerobic conditions at 37°C. The reaction results for a rapid urease activity were determined after 3 hours by a change in the initial color of the medium. The reactions were accounted after 7—24 hours to detect activity urease. It was found that 100% of A. nosocomialis strains and 18.82% of A. baumannii strains had urease activity. At the same time, high activity urease was found only in one strain of A. baumannii (1.17%) and in all strains of A. nosocomialis. Therefore, the presence of high urease activity is of taxonomic importance for the species A. nosocomialis as the marker of distinguishing this species from other bacteria species of the Ab group. The sensitivity and specificity of the identifying A. nosocomialis strains by suggested approach compared to the studied strains identification by MALDI-TOF MS were 100 and 98.83% (x2 = 103.2; p < 0.0001), respectively.E. P. SivolodskiiE. V. ZuevaSankt-Peterburg : NIIÈM imeni Pasteraarticleacinetobacter nosocomialisacinetobacter baumanniibacterial ureaserapid activity ureaseglucose utilizationInfectious and parasitic diseasesRC109-216RUInfekciâ i Immunitet, Vol 11, Iss 3, Pp 591-596 (2021)
institution	DOAJ
collection	DOAJ
language	RU
topic	acinetobacter nosocomialis acinetobacter baumannii bacterial urease rapid activity urease glucose utilization Infectious and parasitic diseases RC109-216
spellingShingle	acinetobacter nosocomialis acinetobacter baumannii bacterial urease rapid activity urease glucose utilization Infectious and parasitic diseases RC109-216 E. P. Sivolodskii E. V. Zueva Method for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria
description	The study's purpose was the developing of the method for Acinetobacter nosocomialis bacteria identification by the totality of phenotypic characteristics of the Acinetobacter baumannii (Ab) group based on the urease activity trait.The research's objects were clinical strains of the Ab group, isolated in 2018—2019 in the bacteriological laboratory of Kirov Military Medical Academy (St. Petersburg), of which were A. baumannii (n = 85), A. nosocomialis (n = 12), A. pittii (n = 10). In addition, 9 strains of A. nosocomialis were isolated from water samples from the Neva River and 2 strains of A. calcoaceticus from the cyanobacterial mats. The belonging of the strains to Ab group was determined by the combination of the metabolic and physiological characteristics of the taxonomic tests for this group. The species identification of the studied strains was determined by the matrix-associated laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). The urease activity of bacteria was determined using the microvolume method. We used a medium with urea in the following composition (g/l): Na2HPO4 — 1.1; KH2PO4 — 1.1; NaCI — 5.0; 40% alkaline solution phenol red — 5 ml; urea — 10—15; distilled water — 1 l. The ingredients, without urea, were dissolved in distilled water, dispensed to vials, sterilized for 20 minutes at 121°C, urea was added in the calculated amount. The medium with urea was added in 0.1 ml to wells of the plate, then inoculated the daily agar culture of the studied and control strains by full loop (2 mm in diameter) and incubated under aerobic conditions at 37°C. The reaction results for a rapid urease activity were determined after 3 hours by a change in the initial color of the medium. The reactions were accounted after 7—24 hours to detect activity urease. It was found that 100% of A. nosocomialis strains and 18.82% of A. baumannii strains had urease activity. At the same time, high activity urease was found only in one strain of A. baumannii (1.17%) and in all strains of A. nosocomialis. Therefore, the presence of high urease activity is of taxonomic importance for the species A. nosocomialis as the marker of distinguishing this species from other bacteria species of the Ab group. The sensitivity and specificity of the identifying A. nosocomialis strains by suggested approach compared to the studied strains identification by MALDI-TOF MS were 100 and 98.83% (x2 = 103.2; p < 0.0001), respectively.
format	article
author	E. P. Sivolodskii E. V. Zueva
author_facet	E. P. Sivolodskii E. V. Zueva
author_sort	E. P. Sivolodskii
title	Method for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria
title_short	Method for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria
title_full	Method for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria
title_fullStr	Method for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria
title_full_unstemmed	Method for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria
title_sort	method for phenotypic identification of <i>acinetobacter nosocomialis</i> bacteria
publisher	Sankt-Peterburg : NIIÈM imeni Pastera
publishDate	2021
url	https://doaj.org/article/01488526aba4406788a953b529e808d6
work_keys_str_mv	AT epsivolodskii methodforphenotypicidentificationofiacinetobacternosocomialisibacteria AT evzueva methodforphenotypicidentificationofiacinetobacternosocomialisibacteria
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Method for phenotypic identification of <i>Acinetobacter nosocomialis</i> bacteria

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