RPB0356

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
F ATATGGTCCTGAAGCAAGTGCATTTACGAA 30 10 μM 40 59.66 9254.09 \
R TGAGCATTATTGACCTGAATCAGCGTTGTC 30 10 μM 43.33 60.52 9212.04 \

Gene Description

Target Gene GenBank ID
nuc gene DQ507382.1

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
the isothermal amplification CRISPR\Cas12a platform is a useful tool for the rapid and sensitive detection of S. aureus in food. RPA-CRISPR\Cas12a Primer Premier 5.0 15 min 37.7 °C CRISPR\Cas12a 5.78 fg\µL of genomic DNA and 67 CFU\mL of S. aureus \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 Isothermal Amplification and CRISPR\Cas12a-System-Based Assay for Rapid, Sensitive and Visual Detection of Staphylococcus aureus Danhong Xu,Haijuan Zeng,Wenhui Wu,Hua Liu,Jinbin Wang Foods 38137236 10.3390/foods12244432

Isothermal Amplification and CRISPR\Cas12a-System-Based Assay for Rapid, Sensitive and Visual Detection of Staphylococcus aureus

Author(s):

Danhong Xu,Haijuan Zeng,Wenhui Wu,Hua Liu,Jinbin Wang

Journal:

Foods

Year:

2023

Abstract:

Staphylococcus aureus exists widely in the natural environment and is one of the main food-borne pathogenic microorganisms causing human bacteremia. For safe food management, a rapid, high-specificity, sensitive method for the detection of S. aureus should be developed. In this study, a platform for detecting S. aureus (nuc gene) based on isothermal amplification (loop-mediated isothermal amplification-LAMP, recombinase polymerase amplification-RPA) and the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas12a) proteins system (LAMP, RPA-CRISPR/Cas12a) was proposed. In this study, the LAMP, RPA-CRISPR/Cas12a detection platform and immunochromatographic test strip (ICS) were combined to achieve a low-cost, simple and visualized detection of S. aureus. The limit of visual detection was 57.8 fg/µL of nuc DNA and 6.7 × 102 CFU/mL of bacteria. Moreover, the platform could be combined with fluorescence detection, namely LAMP, RPA-CRISPR/Cas12a-flu, to establish a rapid and highly sensitive method for the detection of S. aureus. The limit of fluorescence detection was 5.78 fg/µL of genomic DNA and 67 CFU/mL of S. aureus. In addition, this detection platform can detect S. aureus in dairy products, and the detection time was ~40 min. Consequently, the isothermal amplification CRISPR/Cas12a platform is a useful tool for the rapid and sensitive detection of S. aureus in food.