An extraction-free and one-pot two-temperature CRISPR/Cas12b system for visual detection of Group B <i>Streptococcus</i> by integrating with RPA.

ABSTRACT: Group B Streptococcus (GBS) is a major cause of neonatal infections, and rapid detection is essential for timely clinical intervention. In this study, we developed an extraction-free, one-pot CRISPR/Cas12b assay for visual detection of GBS by combining with isothermal amplification, including loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA). The results showed that LAMP-CRISPR/Cas12b outperformed RPA-CRISPR/Cas12b system across all template concentrations, especially in low-copy template (30 and 10 copies/test) detection. To enhance the detection performance of RPA-CRISPR/Cas12b, we introduced a two-temperature protocol, with RPA reaction at 39°C followed by Cas12b activation at 62°C. Through the two-temperature approach, the detection rat

SUBMITTER: Zhao C 

PROVIDER: S-EPMC12506078 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature