RPB0543

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
N. meningitidis Neisseria meningitidis,personal::Sara E. Branham M1027,"Diplokokkus intracellularis meningitidis","Micrococcus meningitidis","Neisseria weichselbaumii","Micrococcus intracellularis","Micrococcus meningitidis cerebrospinalis" 487 Neisseriales Neisseriaceae Neisseria Neisseria meningitidis Bacteria

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 GTCAGGATAAATGGATTGCTCAAGGTTA 28 \ 39.29 56.36 8691.73 \
R CGCATTCGACACATACAATACATCTTTA 28 \ 35.71 54.63 8475.61 \

Gene Description

Target Gene GenBank ID
ctrA gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
This approach could contribute to a new generation of multiplexed field friendly biosensors for improving the diagnostic precision for infectious diseases and their applicability in low-resources settings. RPA PrimerBLAST 20 min 37 °C \ 38 copies\VFMassay \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2018 A vertical flow paper-microarray assay with isothermal DNA amplification for detection of Neisseria meningitidis Lourdes Rivas,Philippa Reuterswärd,Reza Rasti,Björn Herrmann,Andreas Mårtensson,Tobias Alfvén,Jesper Gantelius,Helene Andersson-Svahn Talanta 29567164 10.1016/j.talanta.2018.02.070

A vertical flow paper-microarray assay with isothermal DNA amplification for detection of Neisseria meningitidis

Author(s):

Lourdes Rivas,Philippa Reuterswärd,Reza Rasti,Björn Herrmann,Andreas Mårtensson,Tobias Alfvén,Jesper Gantelius,Helene Andersson-Svahn

Journal:

Talanta

Year:

2018

Abstract:

Paper-based biosensors offer a promising technology to be used at the point of care, enabled by good performance, convenience and low-cost. In this article, we describe a colorimetric vertical-flow DNA microarray (DNA-VFM) that takes advantage of the screening capability of DNA microarrays in a paper format together with isothermal amplification by means of Recombinase Polymerase Amplification (RPA). Different assay parameters such as hybridization buffer, flow rate, printing buffer and capture probe concentration were optimized. A limit of detection (LOD) of 4.4 nM was achieved as determined by tabletop scanning. The DNA-VFM was applied as a proof of concept for detection of Neisseria meningitidis, a primary cause of bacterial meningitis. The LOD was determined to be between 38 and 2.1 × 106 copies/VFMassay, depending on the choice of DNA capture probes. The presented approach provides multiplex capabilities of DNA microarrays in a paper-based format for future point-of-care applications.