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实时聚合酶链反应在孕妇生殖道检测无乳链球菌中的应用。

Performance of RT-PCR in the detection of Streptococcus agalactiae in the anogenital tract of pregnant women.

机构信息

UNISUL, Universidade do Sul de Santa Catarina, Dehon Tubarão, Brazil.

出版信息

Arch Gynecol Obstet. 2012 Dec;286(6):1437-42. doi: 10.1007/s00404-012-2498-9. Epub 2012 Aug 8.

DOI:10.1007/s00404-012-2498-9
PMID:22872028
Abstract

INTRODUCTION

Infection with Group B Streptococcus (GBS) is the most frequent in the first weeks of life of a newborn. The identification of pregnant women with GBS colonization may reduce neonatal infection.

METHODS

This cross-sectional study evaluated the performance of real-time polymerase chain reaction (RT-PCR) to detect GBS colonization in the anogenital tract of pregnant women. Anogenital swabs were collected from 266 pregnant women from December 2010 to August 2011. GBS was detected using culture (gold standard) and RT-PCR to determine sip gene expression. The presence of DNA was confirmed using betaglobin amplification, and the guanidine technique was used for DNA extraction. When results were discordant, the test was repeated using conventional PCR. The results were evaluated to determine sensitivity, specificity, positive and negative predictive values and accuracy.

RESULTS

Of the 266 samples collected, 254 were adequate for analysis. Prevalence was 28.7% using the gold standard criterion and 38.2% using RT-PCR. The comparison of RT-PCR with culture revealed a sensitivity of 89% (95% CI 0.81-0.96), specificity of 82% (95% CI 0.76-0.87), positive predictive value of 67% (95% CI 0.57-0.76) and negative predictive value of 94% (95% CI 0.91-0.99).

CONCLUSION

Further studies using other DNA extraction techniques, targeting other GBS genes and using sample enhancement before RT-PCR should be conducted to determine whether the sensitivity and specificity recommended by the CDC may be reached using the same thermal cycler.

摘要

简介

B 群链球菌(GBS)感染是新生儿生命最初几周最常见的感染。识别具有 GBS 定植的孕妇可降低新生儿感染的风险。

方法

本横断面研究评估了实时聚合酶链反应(RT-PCR)检测孕妇肛门生殖器部位 GBS 定植的性能。采集了 2010 年 12 月至 2011 年 8 月 266 名孕妇的肛门生殖器拭子。使用培养(金标准)和 RT-PCR 检测 GBS,以确定 sip 基因表达。使用β珠蛋白扩增证实 DNA 的存在,并使用胍盐技术进行 DNA 提取。当结果不一致时,使用常规 PCR 重复检测。评估结果以确定敏感性、特异性、阳性和阴性预测值和准确性。

结果

在采集的 266 个样本中,有 254 个样本适合分析。使用金标准标准,患病率为 28.7%,使用 RT-PCR 为 38.2%。RT-PCR 与培养的比较显示敏感性为 89%(95%CI 0.81-0.96),特异性为 82%(95%CI 0.76-0.87),阳性预测值为 67%(95%CI 0.57-0.76),阴性预测值为 94%(95%CI 0.91-0.99)。

结论

应使用其他 DNA 提取技术、针对其他 GBS 基因并在 RT-PCR 前进行样本增强,进一步开展研究,以确定使用相同的热循环仪是否可以达到 CDC 推荐的敏感性和特异性。

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