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Fast, portable and sensitive detection of group B streptococcus DNA using one-pot MIRA-CRISPR system with suboptimal
Weigang Zheng1, Huamei Tang2, Benchen Ye3
1CAS Key Laboratory of Biomedical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Science (CAS), Suzhou, China; Zhengzhou Institute of Biomedical Engineering and Technology, Zhengzhou, China.
A new one-pot diagnostic system accurately detects Group B Streptococcus (GBS) in pregnant women, preventing infant infections. This rapid, extraction-free method enables point-of-care testing for improved maternal and infant health outcomes.
Area of Science:
- Microbiology
- Molecular Diagnostics
- Biotechnology
Background:
- Group B Streptococcus (GBS) poses a significant health risk to infants through vertical transmission during delivery.
- Accurate and rapid prenatal GBS diagnosis is crucial for preventing neonatal infections.
Purpose of the Study:
- To develop an efficient, extraction-free diagnostic assay for Group B Streptococcus (GBS).
- To create a portable system for point-of-care or home-based GBS detection.
Main Methods:
- Developed a one-pot nucleic acid isothermal amplification and CRISPR-Cas12a system for GBS detection.
- Utilized suboptimal protospacer adjacent motifs to minimize handling and contamination.
- Validated the assay using fluorescent techniques and lateral flow assay strips.
Main Results:
- The assay demonstrated high specificity, accuracy, and sensitivity.
- Achieved a limit of detection of 32 copies per reaction (0.64 copies/μL).
- A portable device integrated with the system showed excellent performance in clinical sample detection.
Conclusions:
- The developed one-pot system offers a rapid, sensitive, and specific method for GBS detection.
- The portable device facilitates GBS testing outside traditional laboratory settings, enabling point-of-care and home-self testing.
- This technology has the potential to significantly improve prenatal screening and reduce infant GBS infections.
