RPB0192

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 CTCAAAGCTAGAACTTTGCTTCACTATAAGTATTC 35 \ 34.3 56.27 10664.01 \
R CCCAAGGGCAAAGCGACTTTGTATTCGTCATTGGCGGATCAAACG 45 \ 51.1 70.87 13870.05 \
P TGTTAATTGAACAAGTGTACAGAGCATT-T(FAM)-A-dSpacer-GA-T(BHQ-1)-TATGCGTGGAG 44 \ 37.5 61.89  12429.14 \

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(%)
\ RPA \ 30 25-42 Lateral flow biosensor (LFB) detection assay 10copies μl−1 \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2017 Self-powered integrated microfluidic point-of-care low-cost enabling (SIMPLE) chip Erh-Chia Yeh,Chi-Cheng Fu, Lucy Hu, Rohan Thakur, Jeffrey Feng, and Luke P. Lee Science Advances 28345028 10.1126/sciadv.1501645

Self-powered integrated microfluidic point-of-care low-cost enabling (SIMPLE) chip

Author(s):

Erh-Chia Yeh,Chi-Cheng Fu, Lucy Hu, Rohan Thakur, Jeffrey Feng, and Luke P. Lee

Journal:

Science Advances

Year:

2017

Abstract:

Portable, low-cost, and quantitative nucleic acid detection is desirable for point-of-care diagnostics; however, current polymerase chain reaction testing often requires time-consuming multiple steps and costly equipment. We report an integrated microfluidic diagnostic device capable of on-site quantitative nucleic acid detection directly from the blood without separate sample preparation steps. First, we prepatterned the amplification initiator [magnesium acetate (MgOAc)] on the chip to enable digital nucleic acid amplification. Second, a simplified sample preparation step is demonstrated, where the plasma is separated autonomously into 224 microwells (100 nl per well) without any hemolysis. Furthermore, self-powered microfluidic pumping without any external pumps, controllers, or power sources is accomplished by an integrated vacuum battery on the chip. This simple chip allows rapid quantitative digital nucleic acid detection directly from human blood samples (10 to 105 copies of methicillin-resistant Staphylococcus aureus DNA per microliter, ~30 min, via isothermal recombinase polymerase amplification). These autonomous, portable, lab-on-chip technologies provide promising foundations for future low-cost molecular diagnostic assays.