RPB0316

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Staphylococcus aureus Staphylococcus aureus, Micrococcus aureus, Staphylococcus pyogenes aureus 1280 Bacillales Staphylococcaceae Staphylococcus Staphylococcus aureus Bacterium

Primer Description

Primer Name Sequence(5'-3') Length(bp) Primer Final Concentration(μM） GC Content（%） Predicted Melting Temperature(℃） Molecular Weight(g/moles) Positions in GenBank accession number
nuc-RPA-F GCATCACAAACAGATAACGGCGTAAATAGAAG 32 10 μM 40.63 59.09 9892.54 \
nuc-RPA-R ACATTAATTTAACCGTATCACCATCAATCGCT 32 10 μM 34.38 57.47 9686.39 \

Gene Description

Target Gene GenBank ID
nuc gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
implicity, rapidity, sensitivity, specificity, and strong anti-interference, the biosensor offers a useful tool for evaluating S. aureus contamination in eggs. RPA-CRISPR\Cas12a \ 30 min 37°C CRISPR\Cas12a 3CFU\mL \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2025 Ultrasensitive "On-Off" Ratiometric Fluorescence Biosensor Based on RPA-CRISPR\Cas12a for Detection of Staphylococcus aureus Wang Guo,Yiqing Guo,Hong Xu,Chen Li,Xinai Zhang,Xiaobo Zou,Zongbao Sun Agricultural and Food Chemistry 39791925 10.1021/acs.jafc.4c12202

Ultrasensitive "On-Off" Ratiometric Fluorescence Biosensor Based on RPA-CRISPR\Cas12a for Detection of Staphylococcus aureus

Author(s):

Wang Guo,Yiqing Guo,Hong Xu,Chen Li,Xinai Zhang,Xiaobo Zou,Zongbao Sun

Journal:

Agricultural and Food Chemistry

Year:

2025

Abstract:

Staphylococcus aureus (S. aureus) is a major pathogenic bacterium responsible for bacterial foodborne diseases, making its rapid, specific, and accurate detection crucial. In this study, we develop a ratiometric biosensor based on the recombinase polymerase amplification-clustered regularly interspaced short palindromic repeats/CRISPR associated protein 12a (RPA-CRISPR/Cas12a) system and Eu-metal-organic framework (Eu-MOF) fluorescent nanomaterials for the high-sensitivity detection of S. aureus, combining with RPA for efficient isothermal amplification, this sensor enhances specificity and sensitivity by utilizing the target activation of CRISPR/Cas12a. The Eu-MOF serves a dual function, providing stable red fluorescence as a reference signal and adsorbing FAM-labeled probes for fluorescence quenching, forming a dual-signal system that significantly reduces background interference. This ratiometric design enables accurate and quantitative detection over a wide range (7.9 × 100 to 7.9 × 108 CFU/mL) with a low detection limit of 3 CFU/mL. Overall, with these merits of simplicity, rapid response, high sensitivity, and specificity, this dual-signal biosensor offers a promising method for accurately evaluating S. aureus contamination in food under complex substrate conditions.