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CRISPR-based sensing platform for the Group B streptococcus screening in pregnant women
Tianming Li1, Shiying Chen1, Xiaoying Chen1
1Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Insights
A new portable CRISPR/Cas13a-based device, PalmCS, enables rapid, on-site detection of Group B Streptococcus (GBS). This technology offers a sensitive and specific alternative for prenatal screening, reducing neonatal infection risk.
Area of Science:
- Molecular Diagnostics
- CRISPR-based Detection
- Point-of-Care Testing
Background:
- Group B Streptococcus (GBS) poses a significant risk for perinatal infections.
- Current GBS screening methods like culture are slow, while molecular tests require specialized equipment.
- Existing point-of-care tests are costly and underutilized.
Purpose of the Study:
- To develop a portable CRISPR/Cas13a-based device for rapid, on-site GBS detection.
- To create an efficient nucleic acid extraction and amplification system for GBS screening.
- To provide a cost-effective and accessible diagnostic tool for GBS.
Main Methods:
- Development of a palm-sized CRISPR detection platform (PalmCS) integrating nucleic acid extraction, RPA amplification, and CRISPR/Cas13a detection.
- Optimization of reagents and conditions for a one-pot CRISPR/Cas13a-RPA assay.
- Validation of the PalmCS platform using clinical samples.
Main Results:
- PalmCS achieved rapid nucleic acid extraction in 5 minutes at room temperature.
- The optimized one-pot CRISPR/Cas13a-RPA method demonstrated high efficiency.
- Clinical validation showed a limit of detection of 20 copies/reaction, 97.5% sensitivity, and 100% specificity.
Conclusions:
- The developed PalmCS platform offers a rapid, sensitive, and specific method for GBS detection.
- This technology is suitable for on-site prenatal screening, especially in resource-limited settings.
- On-site GBS testing can guide timely antibiotic administration, significantly reducing neonatal infection risk.
Background:
Group B Streptococcus (GBS) is a major cause of perinatal infections. Prenatal screening is critical to prevent maternal and neonatal GBS infections, reduce adverse outcomes, and guide clinical interventions. While bacterial culture is the gold standard, it is time-consuming and delays decision-making. Rapid molecular tests like PCR are sensitive and specific but require expensive equipment and skilled personnel. Point-of-care tests (e.g., Xpert GBS LB) offer speed and sensitivity but remain costly and underutilized. This study aims to develop a portable CRISPR/Cas13a-based device for rapid, on-site GBS detection.
Results:
A palm-sized CRISPR detection platform, PalmCS, was developed. PalmCS includes two key components: an integrated tube for nucleic acid extraction, gene amplification, and CRISPR-based reaction, and a multifunctional device for thermal regulation, fluorescence detection, and automatic result interpretation. The sealed plug needle valve controls fluid flow for sample input and result output. By optimizing crRNA selection, Cas13a/crRNA ribonucleoprotein (RNP) complex concentrations, and isothermal recombinase polymerase amplification primers, we established a one-pot CRISPR/Cas13a-RPA method for GBS detection. Results showed that 3 M guanidinium salt combined with 5 % Tween 20 achieved the highest extraction efficiency. The system extracted nucleic acids from samples in 5 min at room temperature, demonstrating its potential for rapid diagnostics. PalmCS was validated using 40 clinical samples, achieving a limit of detection (LOD) of 20 copies/reaction, 97.5 % sensitivity, and 100 % specificity.
Significance And Novelty:
This study used a one-step nucleic acid extraction combined with a closed one-pot RPA-CRISPR reaction for rapid, sensitive, and specific GBS detection. This provides a new platform for prenatal screening in pregnant women. In resource-limited settings with emergency deliveries, PalmCS enables on-site GBS testing, allowing antibiotic administration during delivery for positive cases to significantly reduce neonatal infection risk.
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