Advancement in Staphylococcus aureus Detection Using a RPA-CRISPR-Cas12a Fluorescent Assay Technology

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M. Zeinulin

S.Seifullin Kazakh Agrotechnical Research University, Astana, Kazakhstan

M. Amanzholova

National Center for Biotechnology,Astana,010000, Kazakhstan

A. Shaizadinova

al-Farabi Kazakh National University, Almaty, Kazakhstan

S. Abeldenov

S.Seifullin Kazakh Agrotechnical Research University, Astana, Kazakhstan


This study explores the utility of CRISPR/Cas-based diagnostics for the precise identification of Staphylococcus aureus (S. aureus), particularly when integrated with recombinase polymerase amplification (RPA). S. aureus, a versatile Gram-positive bacterium, presents significant challenges due to its antibiotic resistance and its capacity to cause a variety of infections. While existing diagnostic methods provide accuracy, they often involve time-consuming procedures and require specialized equipment. Our research introduces a promising approach that leverages the specificity of CRISPR/Cas technology to target genetic sequences, enabling the precise detection of S. aureus. Through the synergistic combination of CRISPR/Cas and RPA, we expedite the detection process, allowing for rapid and highly sensitive identification of the pathogen's genetic material. Notably, our investigation demonstrates excellent results for the gsa and arcC genes, affirming the potential of this method as a dependable and highly sensitive molecular diagnostic tool. When compared to traditional diagnostic methods such as microbiology and PCR, our approach offers advantages in terms of speed and sensitivity. Additionally, the inclusion of RPA as a pre-amplification step enhances the accuracy of nucleic acid detection. While our study represents an initial step in technological advancement, it highlights the potential of CRISPR/Cas-based diagnostics to revolutionize disease detection and management. The RPA-CRISPR/Cas has the potential for detecting a wide range of diseases and pathogens, with broad applications in both medical and scientific fields.


Staphylococcus aureus, diagnostic, CRISPR/Cas, RPA, Cas12a

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