Do NAAT-Based Methods Increase the Diagnostic Sensitivity of <i>Streptococcus agalactiae</i> 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 <i>Streptococcus</i> (GBS) detection by nucleic acid amplification techni...
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MDPI AG
2023-02-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/13/5/863 |
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| author | Agnieszka Sroka-Oleksiak Wojciech Pabian Joanna Sobońska Kamil Drożdż Tomasz Bogiel Monika Brzychczy-Włoch |
| author_facet | Agnieszka Sroka-Oleksiak Wojciech Pabian Joanna Sobońska Kamil Drożdż Tomasz Bogiel Monika Brzychczy-Włoch |
| author_sort | Agnieszka Sroka-Oleksiak |
| collection | DOAJ |
| 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 <i>Streptococcus</i> (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 <i>16S rRNA</i>, <i>atr</i> and <i>cfb</i> 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 <i>atr</i> gene primers, as compared to <i>cfb</i> and <i>16S rRNA</i> 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 <i>cfb</i> gene, the use of an additional gene to ensure the appropriate results should be considered. |
| first_indexed | 2024-03-11T07:27:56Z |
| format | Article |
| id | doaj.art-c45f74cecb3a48e187131a3bf500186a |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-11T07:27:56Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-c45f74cecb3a48e187131a3bf500186a2023-11-17T07:29:14ZengMDPI AGDiagnostics2075-44182023-02-0113586310.3390/diagnostics13050863Do NAAT-Based Methods Increase the Diagnostic Sensitivity of <i>Streptococcus agalactiae</i> Carriage Detection in Pregnant Women?Agnieszka Sroka-Oleksiak0Wojciech Pabian1Joanna Sobońska2Kamil Drożdż3Tomasz Bogiel4Monika Brzychczy-Włoch5Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 31-121 Krakow, PolandClinical Department of Gynecological Endocrinology and Gynecology, Jagiellonian University Medical College, 31-501 Krakow, PolandDepartment of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 31-121 Krakow, PolandDepartment of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 31-121 Krakow, PolandDepartment of Microbiology, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, 85-094 Bydgoszcz, PolandDepartment of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 31-121 Krakow, PolandThe 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 <i>Streptococcus</i> (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 <i>16S rRNA</i>, <i>atr</i> and <i>cfb</i> 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 <i>atr</i> gene primers, as compared to <i>cfb</i> and <i>16S rRNA</i> 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 <i>cfb</i> gene, the use of an additional gene to ensure the appropriate results should be considered.https://www.mdpi.com/2075-4418/13/5/863GBSPCRpregnant womenprimersreal-time PCR<i>Streptococcus agalactiae</i> |
| spellingShingle | Agnieszka Sroka-Oleksiak Wojciech Pabian Joanna Sobońska Kamil Drożdż Tomasz Bogiel Monika Brzychczy-Włoch Do NAAT-Based Methods Increase the Diagnostic Sensitivity of <i>Streptococcus agalactiae</i> Carriage Detection in Pregnant Women? Diagnostics GBS PCR pregnant women primers real-time PCR <i>Streptococcus agalactiae</i> |
| title | Do NAAT-Based Methods Increase the Diagnostic Sensitivity of <i>Streptococcus agalactiae</i> Carriage Detection in Pregnant Women? |
| title_full | Do NAAT-Based Methods Increase the Diagnostic Sensitivity of <i>Streptococcus agalactiae</i> Carriage Detection in Pregnant Women? |
| title_fullStr | Do NAAT-Based Methods Increase the Diagnostic Sensitivity of <i>Streptococcus agalactiae</i> Carriage Detection in Pregnant Women? |
| title_full_unstemmed | Do NAAT-Based Methods Increase the Diagnostic Sensitivity of <i>Streptococcus agalactiae</i> Carriage Detection in Pregnant Women? |
| title_short | Do NAAT-Based Methods Increase the Diagnostic Sensitivity of <i>Streptococcus agalactiae</i> Carriage Detection in Pregnant Women? |
| title_sort | do naat based methods increase the diagnostic sensitivity of i streptococcus agalactiae i carriage detection in pregnant women |
| topic | GBS PCR pregnant women primers real-time PCR <i>Streptococcus agalactiae</i> |
| url | https://www.mdpi.com/2075-4418/13/5/863 |
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