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Do NAAT-Based Methods Increase the Diagnostic Sensitivity of Streptococcus agalactiae Carriage Detection in Pregnant Women?

The aim of the study was to evaluate particular polymerase chain reaction primers targeting selected representative genes and the influence of a preincubation step in a selective broth on the sensitivity of group B Streptococcus (GBS) detection by nucleic acid amplification techniques (NAAT). Resear...

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Autores principales: Sroka-Oleksiak, Agnieszka, Pabian, Wojciech, Sobońska, Joanna, Drożdż, Kamil, Bogiel, Tomasz, Brzychczy-Włoch, Monika
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10001255/
https://www.ncbi.nlm.nih.gov/pubmed/36900007
http://dx.doi.org/10.3390/diagnostics13050863
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author Sroka-Oleksiak, Agnieszka
Pabian, Wojciech
Sobońska, Joanna
Drożdż, Kamil
Bogiel, Tomasz
Brzychczy-Włoch, Monika
author_facet Sroka-Oleksiak, Agnieszka
Pabian, Wojciech
Sobońska, Joanna
Drożdż, Kamil
Bogiel, Tomasz
Brzychczy-Włoch, Monika
author_sort Sroka-Oleksiak, Agnieszka
collection PubMed
description The aim of the study was to evaluate particular polymerase chain reaction primers targeting selected representative genes and the influence of a preincubation step in a selective broth on the sensitivity of group B Streptococcus (GBS) detection by nucleic acid amplification techniques (NAAT). Research samples were vaginal and rectal swabs collected in duplicate from 97 pregnant women. They were used for enrichment broth culture-based diagnostics, bacterial DNA isolation, and amplification, using primers based on species-specific 16S rRNA, atr and cfb genes. To assess the sensitivity of GBS detection, additional isolation of samples preincubated in Todd-Hewitt broth with colistin and nalidixic acid was performed and then subjected to amplification again. The introduction of the preincubation step increased the sensitivity of GBS detection by about 33–63%. Moreover, NAAT made it possible to identify GBS DNA in an additional six samples that were negative in culture. The highest number of true positive results compared to the culture was obtained with the atr gene primers, as compared to cfb and 16S rRNA primers. Isolation of bacterial DNA after preincubation in enrichment broth significantly increases the sensitivity of NAAT-based methods applied for the detection of GBS from vaginal and rectal swabs. In the case of the cfb gene, the use of an additional gene to ensure the appropriate results should be considered.
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spelling pubmed-100012552023-03-11 Do NAAT-Based Methods Increase the Diagnostic Sensitivity of Streptococcus agalactiae Carriage Detection in Pregnant Women? Sroka-Oleksiak, Agnieszka Pabian, Wojciech Sobońska, Joanna Drożdż, Kamil Bogiel, Tomasz Brzychczy-Włoch, Monika Diagnostics (Basel) Article The aim of the study was to evaluate particular polymerase chain reaction primers targeting selected representative genes and the influence of a preincubation step in a selective broth on the sensitivity of group B Streptococcus (GBS) detection by nucleic acid amplification techniques (NAAT). Research samples were vaginal and rectal swabs collected in duplicate from 97 pregnant women. They were used for enrichment broth culture-based diagnostics, bacterial DNA isolation, and amplification, using primers based on species-specific 16S rRNA, atr and cfb genes. To assess the sensitivity of GBS detection, additional isolation of samples preincubated in Todd-Hewitt broth with colistin and nalidixic acid was performed and then subjected to amplification again. The introduction of the preincubation step increased the sensitivity of GBS detection by about 33–63%. Moreover, NAAT made it possible to identify GBS DNA in an additional six samples that were negative in culture. The highest number of true positive results compared to the culture was obtained with the atr gene primers, as compared to cfb and 16S rRNA primers. Isolation of bacterial DNA after preincubation in enrichment broth significantly increases the sensitivity of NAAT-based methods applied for the detection of GBS from vaginal and rectal swabs. In the case of the cfb gene, the use of an additional gene to ensure the appropriate results should be considered. MDPI 2023-02-23 /pmc/articles/PMC10001255/ /pubmed/36900007 http://dx.doi.org/10.3390/diagnostics13050863 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sroka-Oleksiak, Agnieszka
Pabian, Wojciech
Sobońska, Joanna
Drożdż, Kamil
Bogiel, Tomasz
Brzychczy-Włoch, Monika
Do NAAT-Based Methods Increase the Diagnostic Sensitivity of Streptococcus agalactiae Carriage Detection in Pregnant Women?
title Do NAAT-Based Methods Increase the Diagnostic Sensitivity of Streptococcus agalactiae Carriage Detection in Pregnant Women?
title_full Do NAAT-Based Methods Increase the Diagnostic Sensitivity of Streptococcus agalactiae Carriage Detection in Pregnant Women?
title_fullStr Do NAAT-Based Methods Increase the Diagnostic Sensitivity of Streptococcus agalactiae Carriage Detection in Pregnant Women?
title_full_unstemmed Do NAAT-Based Methods Increase the Diagnostic Sensitivity of Streptococcus agalactiae Carriage Detection in Pregnant Women?
title_short Do NAAT-Based Methods Increase the Diagnostic Sensitivity of Streptococcus agalactiae Carriage Detection in Pregnant Women?
title_sort do naat-based methods increase the diagnostic sensitivity of streptococcus agalactiae carriage detection in pregnant women?
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10001255/
https://www.ncbi.nlm.nih.gov/pubmed/36900007
http://dx.doi.org/10.3390/diagnostics13050863
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