Sroka-Oleksiak Agnieszka, Pabian Wojciech, Sobońska Joanna, Drożdż Kamil, Bogiel Tomasz, Brzychczy-Włoch Monika
Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 31-121 Krakow, Poland.
Clinical Department of Gynecological Endocrinology and Gynecology, Jagiellonian University Medical College, 31-501 Krakow, Poland.
Diagnostics (Basel). 2023 Feb 23;13(5):863. doi: 10.3390/diagnostics13050863.
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 (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 , and 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 gene primers, as compared to and 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 gene, the use of an additional gene to ensure the appropriate results should be considered.
本研究的目的是评估针对选定代表性基因的特定聚合酶链反应引物,以及选择性肉汤中的预孵育步骤对核酸扩增技术(NAAT)检测B族链球菌(GBS)敏感性的影响。研究样本为从97名孕妇中重复采集的阴道和直肠拭子。这些拭子用于基于富集肉汤培养的诊断、细菌DNA分离和扩增,使用基于种特异性、和基因的引物。为评估GBS检测的敏感性,对在含有黏菌素和萘啶酸的托德-休伊特肉汤中预孵育的样本进行了额外分离,然后再次进行扩增。预孵育步骤的引入使GBS检测的敏感性提高了约33%-63%。此外,NAAT能够在另外六个培养结果为阴性的样本中鉴定出GBS DNA。与和引物相比,使用基因引物获得的真阳性结果数量与培养结果相比最多。在富集肉汤中预孵育后分离细菌DNA可显著提高基于NAAT的方法从阴道和直肠拭子中检测GBS的敏感性。对于基因,应考虑使用额外的基因以确保获得合适的结果。