Streptococcus agalactiae (GBS) is the causative pathogen of puerperal sepsis in pregnant women and pneumonia, sepsis and meningitis in infants. Infection of GBS is responsible for the increased morbidity in pregnant women and the elderly, and bring challenges to clinical diagnosis and treatment. However, culture-based approaches to detect S.agalactiae is time-consuming with limited sensitivity. Besides, real-time quantitative PCR demands expensive instruments with tedious steps. Thus, we aim to establish a new detection method for more accurate and rapid detection of S.agalactiae. The ddPCR primer targeted the CpsE gene showed better amplified efficiency in the reaction. The limit of detection for GBS DNA with ddPCR was able to reach 5 pg/μL. Moreover, no positive amplified signals could be detected in the reactions which served 11 non-GBS strains DNA as templates. Furthermore, the coefficient of variation of this method was 4.5%, indicating excellent repeatability of ddPCR assay. In our study, ddPCR was performed as a rapid detection of S.agalactiae with high sensitivity and specificity. This technique can promote the accuracy of the diagnosis of GBS infection and provide a scientific basis for clinical treatment.

中文翻译：

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用于检测无乳链球菌的液滴数字PCR方法的开发。

无乳链球菌（GBS）是孕妇产后败血症和婴儿肺炎，败血症和脑膜炎的病原体。GBS感染是孕妇和老年人发病率上升的原因，并给临床诊断和治疗带来挑战。但是，基于文化的方法来检测龙舌兰酵母很费时且灵敏度有限。此外，实时定量PCR要求昂贵的仪器，且步骤繁琐。因此，我们旨在建立一种新的检测方法，以更准确，快速地检测龙舌兰。靶向CpsE基因的ddPCR引物在反应中显示出更好的扩增效率。ddPCR对GBS DNA的检出限能够达到5 pg /μL。此外，在以11种非GBS菌株DNA为模板的反应中，没有检测到阳性扩增信号。此外，该方法的变异系数为4.5％，表明ddPCR测定具有极好的可重复性。在我们的研究中，以高灵敏度和特异性快速检测ddagalactiae。该技术可以提高GBS感染诊断的准确性，为临床治疗提供科学依据。