Fluorescence polarization system for rapid COVID-19 diagnosis

Lee, Chang Yeol; Degani, Ismail; Cheong, Jiyong; Lee, Jae Hyun; Choi, Hyun‐Jung; Cheon, Jinwoo; Lee, Hakho

doi:10.1016/j.bios.2021.113049

Cited by 50 publications

(44 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the amplification can be adapted to be carried out at room temperature with optimization, it is necessary to integrate the sample treatment and detection reaction into a single reaction. Currently, the fluorescence read-out is dependent on a lab-set fluorimeter, and it is necessary to integrate a portable fluorescence reader as used in other POCT systems [ 38 ]. Additionally, although the LFA read-out can show public-friendly, easy-to-read results without further analysis, the whole reaction from sample preparation to detection is handled in the lab.…”

Section: Discussionmentioning

confidence: 99%

“…There are several other SARS-CoV-2 detection approaches that have been reported recently that work as a single-step reaction, integrated portable devices for the purpose of POCT, such as CRISPR optical detection of anisotropy (CODA), CRISPR-assisted detection (CASdetec), personal glucose meter–CRISPR assay (PGM-CRISPR), an integrated portable droplet magnetofluidic (DM) device, and CRISPR-fluorescence detection system (CRISPR-FDS) [ 38 , 39 , 40 , 41 , 42 , 43 , 44 ]. While these approaches are already functional POCT devices, they are capable of detecting SARS-CoV-2 only.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Detection of Infectious Viruses Using CRISPR-Cas12-Based Assay

et al. 2021

View full text Add to dashboard Cite

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease-19 (COVID-19), has severely influenced public health and economics. For the detection of SARS-CoV-2, clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein (Cas)-based assays have been emerged because of their simplicity, sensitivity, specificity, and wide applicability. Herein, we have developed a CRISPR-Cas12-based assay for the detection of SARS-CoV-2. In the assay, the target amplicons are produced by isothermal reverse transcription recombinase polymerase amplification (RT-RPA) and recognized by a CRISPR-Cas12a/guide RNA (gRNA) complex that is coupled with the collateral cleavage activity of fluorophore-tagged probes, allowing either a fluorescent measurement or naked-eye detection on a lateral flow paper strip. This assay enables the sensitive detection of SARS-CoV-2 at a low concentration of 10 copies per sample. Moreover, the reliability of the method is verified by using nasal swabs and sputum of COVID-19 patients. We also proved that the current assay can be applied to other viruses, such as Middle East respiratory syndrome coronavirus (MERS-CoV) and severe acute respiratory syndrome coronavirus (SARS-CoV), with no major changes to the basic scheme of testing. It is anticipated that the CRISPR-Cas12-based assay has the potential to serve as a point-of-care testing (POCT) tool for a wide range of infectious viruses.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Detection of Infectious Viruses Using CRISPR-Cas12-Based Assay

et al. 2021

View full text Add to dashboard Cite

show abstract

“…In a total of 50 specimens, viral RNAs were isolated from 38 pharyngeal swabs using each 200 µL of a viral transport media with the KingFisher Flex system (ThermoFisher Scientific Inc., Worcester, MA, USA), an automated nucleic acid extraction system, following the manufacturer’s guidelines protocol (Lin et al 2021 ). In addition, we isolated viral RNAs using the Advansure™ E3 system (LG Chem, Ltd., Seoul, Korea), an automated nucleic acid extraction system, and 200 µL each of 12 viral transport media obtained from sputum specimens according to the manufacturer’s guideline (Lee et al 2021 ). The viral RNAs isolated by each system were dissolved in 50 µL of RNase-free water.…”

Section: Methodsmentioning

confidence: 99%

Diagnostic evaluation of qRT-PCR-based kit and dPCR-based kit for COVID-19

et al. 2021

View full text Add to dashboard Cite

Background Coronavirus disease of 2019 (COVID-19) is well known as a fatal disease, first discovered at Wuhan in China, ranging from mild to death, such as shortness of breath and fever. Early diagnosis of COVID-19 is a crucial point in preventing global prevalence. Objective We aimed to evaluate the diagnostic competency and efficiency with the Allplex™ 2019-nCoV Assay kit and the Dr. PCR 20 K COVID-19 Detection kit, designed based on the qRT-PCR and dPCR technologies, respectively. Methods A total of 30 negative and 20 COVID-19 positive specimens were assigned to the diagnostic test by using different COVID-19 diagnosis kits. Diagnostic accuracy was measured by statistical testing with sensitivity, specificity, and co-efficiency calculations. Results Comparing both diagnostic kits, we confirmed that the diagnostic results of 30 negative and 20 positive cases were the same pre-diagnostic results. The diagnostic statistics test results were perfectly matched with value (1). Cohen’s Kappa coefficient was demonstrated that the given kits in two different ways were “almost perfect” with value (1). In evaluating the detection capability, the dilutional linearity experiments substantiate that the Dr. PCR 20 K COVID-19 Detection kit could detect SARS-CoV-2 viral load at a concentration ten times lower than that of the Allplex™ 2019-nCoV Assay kit. Conclusions In this study, we propose that the dPCR diagnosis using LOAA dPCR could be a powerful method for COVID-19 point-of-care tests requiring immediate diagnosis in a limited time and space through the advantages of relatively low sample concentration and small equipment size compared to conventional qRT-PCR. Supplementary Information The online version contains supplementary material available at 10.1007/s13258-021-01162-4.

show abstract

“…It is of great importance to develop sensitive and accurate SARS-CoV-2 detection methods to promote the epidemic prevention and control of COVID-19. Currently, reverse transcription-polymerase chain reaction (RT-PCR) is the principal method to identify SARS-CoV-2 infection [5] , [6] , [7] . However, RT-PCR requires expensive special equipment, a reverse transcription step and well-trained technicians [8] , [9] , [10] , [11] , [12] .…”

Section: Introductionmentioning

confidence: 99%