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. 2020 Jun 23;20(1):179.
doi: 10.1186/s12866-020-01857-w.

Development of a droplet digital PCR method for detection of Streptococcus agalactiae

Yi-Fan Zeng  1   2   3   4 , Chu-Mao Chen  1   2 , Xiao-Yan Li  5 , Jun-Jiang Chen  1   2   3 , Yan-Ge Wang  1   2   3 , Shi Ouyang  6 , Tian-Xing Ji  7 , Yong Xia  1   2   3 , Xu-Guang Guo  8   9   10   11   12   13
Affiliations

Development of a droplet digital PCR method for detection of Streptococcus agalactiae

Yi-Fan Zeng et al. BMC Microbiol. .

Abstract

Background: 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.

Results: 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.

Conclusions: 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.

Keywords: Droplet digital PCR; Quantitation; Streptococcus agalactiae.

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Conflict of interest statement

The authors declare that there are no competing interests associated with the manuscript.

Figures

Fig. 1
Fig. 1
The primer of the qPCR screening experiment in the present study
Fig. 2
Fig. 2
Sensitivity assay for ddPCR using continuous 10-fold dilutions of DNA templates from GBS ATCC13813
Fig. 3
Fig. 3
The number of events in each amplification concentration
Fig. 4
Fig. 4
The repeatability test of ddPCR using DNA from the GBS ATCC13813 strain
See this image and copyright information in PMC

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