RPB0246

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
\ \ \ 10 μM \ \ \ \
\ \ \ 10 μM \ \ \ \

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(%)
\ RPA-LFIA \ 20 min 38°C LFIA readout system 4.6 × 102CFU\mL \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2025 Dual recombinase polymerase amplification system combined with lateral flow immunoassay for simultaneous detection of Staphylococcus aureus and Vibrio parahaemolyticus Yan Zhang,Xiaofeng Liu,Jiawei Luo,Hua Liu,You Li,Juan Liu,Lemei Zhu,Jinbin Wang,Haijuan Zeng Journal of Pharmaceutical and Biomedical Analysis 39644678 10.1016/j.jpba.2024.116621

Dual recombinase polymerase amplification system combined with lateral flow immunoassay for simultaneous detection of Staphylococcus aureus and Vibrio parahaemolyticus

Author(s):

Yan Zhang,Xiaofeng Liu,Jiawei Luo,Hua Liu,You Li,Juan Liu,Lemei Zhu,Jinbin Wang,Haijuan Zeng

Journal:

Journal of Pharmaceutical and Biomedical Analysis

Year:

2025

Abstract:

Development of a highly sensitive visualization platform for multiplex genetic detection could significantly improve efficiency and reliability of on-site detection of foodborne pathogens. In this study, coupling recombinase polymerase amplification (RPA) with lateral flow immunoassay (LFIA) readout system was proposed for Staphylococcus aureus and Vibrio parahaemolyticus detection. Taking the advantage of the isothermal amplification of RPA, dual primers modified with different labeling groups were designed to realize target signal amplification. LFIA coated with anti-digoxigenin antibody and streptavidin as test line 1 and 2 were designed to detect the two RPA products. The proposed method (dual RPA-LFIA) could realize visual detection using LFIA through rapid RPA amplification within 20 min, exhibiting a lowest detection limit of 4.6 × 102 CFU/mL for Staphylococcus aureus and Vibrio parahaemolyticus. The dual RPA-LFIA is characterized by simultaneous detection of dual targets in one RPA reaction and colorimetric readout through LFIA, thus ensuring high sensitivity and efficiency, and showing great potential to address the on-site detection of foodborne pathogens in the future.