RPB0013

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
SARS-CoV-2 SARS-CoV-2, 2019-nCoV, COVID-19, COVID-19 virus, SARS2, Wuhan coronavirus, Human coronavirus 2019, COVID19, HCoV-19, SARS-2, SARS-CoV4 2697049 Nidovirales Coronaviridae Betacoronavirus Severe acute respiratory syndrome-related coronavirus virus

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 ACACCGGAAGCCAATATGGATCAAGAATCC 30 \ 46.7 62.13 9202.07 \
R ACCCACAGGGTCATTAGCACAAGTTGTAGG 30 \ 50 63.84 9240.07 \
P ATCGTGTTGTCTGTACTGCCGTTGCCACA/i6FAMdT/A/idSp/A/iBHQ1dT/CATCCAAATCCTAAAGG 48 \ 44 68.09 15294.98 \

Gene Description

Target Gene GenBank ID
ORF1ab \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
fast and accurate on-site diagnostic testing for infectious pathogen RPA-POCT \ 20 40 fluorescence(under 488 nm blue light excitation) 400 copies/mL \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 Portable Paper-Based Nucleic Acid Enrichment for Field Testing Junyang Mei, Dandan Wang, Yiheng Zhang, Dan Wu, Jinhui Cui, Mingzhe Gan, Peifeng Liu Advanced Science 36797206 10.1002/advs.202205217

Portable Paper-Based Nucleic Acid Enrichment for Field Testing

Author(s):

Junyang Mei, Dandan Wang, Yiheng Zhang, Dan Wu, Jinhui Cui, Mingzhe Gan, Peifeng Liu

Journal:

Advanced Science

Year:

2023

Abstract:

Point-of-care testing (POCT) can be the method of choice for detecting infectious pathogens; these pathogens are responsible for not only infectious diseases such as COVID-19, but also for certain types of cancers. For example, infections by human papillomavirus (HPV) or Helicobacter pylori (H. pylori) are the main cause of cervical and stomach cancers, respectively. COVID-19 and many cancers are treatable with early diagnoses using POCT. A variety of nucleic acid testing have been developed for use in resource-limited environments. However, questions like unintegrated nucleic acid extraction, open detection systems increase the risk of cross-contamination, and dependence on expensive equipment and alternating current (AC) power supply, significantly limit the application of POCT, especially for on-site testing. In this paper, a simple portable platform is reported capable of rapid sample-to-answer testing within 30 min based on recombinase polymerase amplification (RPA) at a lower temperature, to detect SARS-CoV-2 virus and H. pylori bacteria with a limit of detection as low as 4 × 102 copies mL-1 . The platform used a battery-powered portable reader for on-chip one-pot amplification and fluorescence detection, and can test for multiple (up to four) infectious pathogens simultaneously. This platform can provide an alternative method for fast and reliable on-site diagnostic testing.