RPB0278

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
SA-F TATATGACAGCAACACCACCGAAACAACTTCT 32 600 nM 40.63 60.88 9714.42 \
SA-R CCAGTGTATAACGTTGATTATTAATTTGATCT 32 600 nM 28.13 52.47 9818.46 \

Gene Description

Target Gene GenBank ID
\ \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
\ RAA-CRISPR\Cas13a \ 70 MIN 39°C CRISPR\Cas13a system \ 101Cp/μL \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2024 Multiplex bacteria detection using one-pot CRISPR\Cas13a-based droplet microfluidics Yuting Shang,Gaowa Xing,Jiaxu Lin,Yuxuan Li,Yongning Lin,Shulang Chen,Jin-Ming Lin Biosensors and Bioelectronics 37875060 10.1016/j.bios.2023.115771

Multiplex bacteria detection using one-pot CRISPR\Cas13a-based droplet microfluidics

Author(s):

Yuting Shang,Gaowa Xing,Jiaxu Lin,Yuxuan Li,Yongning Lin,Shulang Chen,Jin-Ming Lin

Journal:

Biosensors and Bioelectronics

Year:

2024

Abstract:

High-throughput detection of bacteria at low levels is critical in public health, food safety, and first response. Herein, for the first time, we present a platform based on droplet microfluidics coupling with the recombinase aided amplification (RAA)-assisted one-pot clustered regularly interspaced short palindromic repeats together with CRISPR-associated proteins 13a (CRISPR/Cas13a) assay, and droplet encoding strategy for accurate and sensitive determination of nucleic acids from various foodborne pathogens. The workflow takes full advantage of CRISPR/Cas13a signal amplification and droplet confinement effects, which enhances the detection sensitivity and enables end-point quantitation. Meanwhile, by varying the color of droplets, the number of bacteria detected at the same time is greatly improved. It possesses the capability to simultaneously detect seven different types of foodborne pathogens. Notably, the system is also applied to real food samples with satisfactory results. Overall, in view of superiorities in high sensitivity, outstanding selectivity, and large-scale multiplexing, the one-pot CRISPR/Cas13a-based droplet microfluidic system could be expanded and universalized for identifying other bacteria.