RPB0036

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
OF TGTAGTTGTGATCAACTCCGCGAACCCATGCT 32 \ 50 66.34 9775.4 \
OR TCTTCATGTTGGTAGTTAGAGAAAGTGTGTCT 32 \ 37.5 58.17 9915.5 \
F TCAGTCAGCTGATGCACAATCGTTTTTAAACG 32 \ 40.6 60.7 9798.44 \
R CTTGGAAGCGACAACAATTAGTTTTTAGGA 30 \ 36.7 56.85 9269.11 \
P AGCCCGTCTTACACCGTGCGGCACAGGCAC/i6FAMdT/idSp/G/iBHQ1dT/ACTGATGTCGTAT/C3 spacer 44 \ 56.5 74.71 14102.17 \

Gene Description

Target Gene GenBank ID
ORF1ab NC_045512.2

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
monitoring of aerosols in a public space SARSCoV-2-in-aerosol monitoring system (SIAMs) Primer-BLAST 30 42 fluorescence 20 copies of viruses per mL \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2022 An ultrasensitive and rapid "sample-to-answer" microsystem for on-site monitoring of SARS-CoV-2 in aerosols using "in situ" tetra-primer recombinase polymerase amplification Shanglin Li, Bao Li, Xinyue Li, Ce Liu, Xiao Qi, Yin Gu, Baobao Lin, Lingli Sun, Lan Chen, Bingqian Han, Jiazhen Guo, Yanyi Huang, Shuangsheng Wu, Lili Ren, Jianbin Wang, Jingwei Bai, Jianxin Ma, Maosheng Yao, Peng Liu Biosensors & Bioelectronics 36327559 10.1016/j.bios.2022.114816

An ultrasensitive and rapid "sample-to-answer" microsystem for on-site monitoring of SARS-CoV-2 in aerosols using "in situ" tetra-primer recombinase polymerase amplification

Author(s):

Shanglin Li, Bao Li, Xinyue Li, Ce Liu, Xiao Qi, Yin Gu, Baobao Lin, Lingli Sun, Lan Chen, Bingqian Han, Jiazhen Guo, Yanyi Huang, Shuangsheng Wu, Lili Ren, Jianbin Wang, Jingwei Bai, Jianxin Ma, Maosheng Yao, Peng Liu

Journal:

Biosensors & Bioelectronics

Year:

2022

Abstract:

Airborne transmissibility of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has highlighted the urgent need for aerosol monitoring of SARS-CoV-2 to prevent sporadic outbreaks of COVID-19. The inadequate sensitivity of conventional methods and the lack of an on-site detection system limited the practical SARS-CoV-2 monitoring of aerosols in public spaces. We have developed a novel SARS-CoV-2-in-aerosol monitoring system (SIAMs) which consists of multiple portable cyclone samplers for collecting aerosols from several venues and a sensitive "sample-to-answer" microsystem employing an integrated cartridge for the analysis of SARS-CoV-2 in aerosols (iCASA) near the sampling site. By seamlessly combining viral RNA extraction based on a chitosan-modified quartz filter and "in situ" tetra-primer recombinase polymerase amplification (tpRPA) into an integrated microfluidic cartridge, iCASA can provide an ultra-high sensitivity of 20 copies/mL, which is nearly one order of magnitude greater than that of the commercial kit, and a short turnaround time of 25 min. By testing various clinical samples of nasopharyngeal swabs, saliva, and exhaled breath condensates obtained from 23 COVID-19 patients, we demonstrate that the positive rate of our system was 3.3 times higher than those of the conventional method. Combining with multiple portable cyclone samplers, we detected 52.2% (12/23) of the aerosol samples, six times higher than that of the commercial kit, collected from the isolation wards of COVID-19 patients, demonstrating the excellent performance of our system for SARS-CoV-2-in-aerosol monitoring. We envision the broad application of our microsystem in aerosol monitoring for fighting the COVID-19 pandemic.