Paper-based netlike rolling circle amplification (NRCA) for ultrasensitive and visual detection of SARS-CoV-2

Song, Yuanlin; Chao, Yuqing; Guo, Yi; Zhang, Fan; Mao, Changqing; Guan, Chaoyang; Chen, Guifang; Feng, Chao

doi:10.1016/j.snb.2022.131460

Cited by 19 publications

(9 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Following the reference calculation method, the equation for the fitted curve was obtained: Y = 0.0229 + 0.1907 × log 10 X (R 2 = 0.9891), and the LOD was calculated as 1.49 aM by using the formula LOD = 3 SD/k, where SD represents the standard deviation of the blank sample and k is the slope of the fitted curve (Figure 2f). 50 Then, we verified the specificity of the mCAD strategy on artificially mutated target sequences, including one-, two-, three-, and five-base mismatches (mT-1, mT-2, mT-3, and mT-5, respectively) and totally unmatched variants (uT). Both PAGE (Figures 2g,h and S7) and fluorescence measurements (Figure 2i) revealed that mCAD was able to successfully discriminate mutated target sequences with three-base mismatches and above but failed to discriminate targets with merely 1-base or 2-base mismatches.…”

Section: ■ Results and Discussionmentioning

confidence: 97%

“…The sample with a concentration of 10 aM showed a notable distinction from the negative control (Figure e). Following the reference calculation method, the equation for the fitted curve was obtained: Y = 0.0229 + 0.1907 × log 10 X ( R 2 = 0.9891), and the LOD was calculated as 1.49 aM by using the formula LOD = 3 SD/ k , where SD represents the standard deviation of the blank sample and k is the slope of the fitted curve (Figure f) …”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Precise Detection of Viral RNA by Programming Multiplex Rolling Circle Amplification and Strand Displacement

Wang,

Ma,

et al. 2023

Anal. Chem.

View full text Add to dashboard Cite

Detection of viral infections (e.g., SARS-CoV-2) with high precision is critical to disease control and treatment. There is an urgent need to develop point-of-care detection methods to complement the gold standard laboratory-based PCR assay with comparable sensitivity and specificity. Herein, we developed a method termed mCAD to achieve ultraspecific point-of-care detection of SARS-CoV-2 RNA while maintaining high sensitivity by programming multiplex rolling circle amplification and toehold-mediated strand displacement reactions. RCA offers sufficient amplification of RNA targets for subsequent detection. Most importantly, a multilayer of detection specificity is implemented into mCAD via sequence-specific hybridization of nucleic acids across serial steps of this protocol to fully eliminate potential false-positive detections. Using mCAD, we demonstrated a highly specific, sensitive, and convenient visual detection of SARS-CoV-2 RNA from both synthetic and clinical samples, exhibiting performance comparable to qPCR. We envision that mCAD will find its broad applications in clinical prospects for nucleic acid detections readily beyond SARS-CoV-2 RNA.

show abstract

Section: ■ Results and Discussionmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Precise Detection of Viral RNA by Programming Multiplex Rolling Circle Amplification and Strand Displacement

Wang,

Ma,

et al. 2023

Anal. Chem.

View full text Add to dashboard Cite

show abstract

“…While most paper-based RCA demonstrated only detection, two articles demonstrated both amplification and detection [ 116 , 117 ]. Both works utilized fluorometric detection with intercalating dyes, including SYBR Green and QuantiFluor dye.…”

Section: Rca With Paper Microfluidicsmentioning

confidence: 99%

“…Both works utilized fluorometric detection with intercalating dyes, including SYBR Green and QuantiFluor dye. In the first publication [ 116 ], test areas were wax printed onto a nitrocellulose membrane, and a blank film was created by printing and drying pullulan solution onto the paper. Next, the RCA reagents, including padlock probe, primer 1, primer 2, T4 DNA ligase, and SYBR Green I, were added to create the bioactive film.…”

Section: Rca With Paper Microfluidicsmentioning

confidence: 99%

Recent Uses of Paper Microfluidics in Isothermal Nucleic Acid Amplification Tests

Reynolds,

Loeffler,

Leigh

et al. 2023

Biosensors

View full text Add to dashboard Cite

Isothermal nucleic acid amplification tests have recently gained popularity over polymerase chain reaction (PCR), as they only require a constant temperature and significantly simplify nucleic acid amplification. Recently, numerous attempts have been made to incorporate paper microfluidics into these isothermal amplification tests. Paper microfluidics (including lateral flow strips) have been used to extract nucleic acids, amplify the target gene, and detect amplified products, all toward automating the process. We investigated the literature from 2020 to the present, i.e., since the onset of the COVID-19 pandemic, during which a significant surge in isothermal amplification tests has been observed. Paper microfluidic detection has been used extensively for recombinase polymerase amplification (RPA) and its related methods, along with loop-mediated isothermal amplification (LAMP) and rolling circle amplification (RCA). Detection was conducted primarily with colorimetric and fluorometric methods, although a few publications demonstrated flow distance- and surface-enhanced Raman spectroscopic (SERS)-based detection. A good number of publications could be found that demonstrated both amplification and detection on paper microfluidic platforms. A small number of publications could be found that showed extraction or all three procedures (i.e., fully integrated systems) on paper microfluidic platforms, necessitating the need for future work.

show abstract

“…Since the emergence of the coronavirus disease 2019 (COVID-19) pandemic, there has been an acute need felt for reliable and applications in biology, science, and medicine due to its simplicity but challenges in sample purification, reaction time, mass production, and shelf storage time. [12,19] Next, the lateral flow assay (LFA) has high speed, specificity, and simplicity. [20,21] Still, it has extremely poor sensitivity as the test is based on detecting the antibody response to an infection and viral antigens.…”

Section: Introductionmentioning

confidence: 99%

Mobile Efficient Diagnostics of Infectious Diseases via On‐Chip RT‐qPCR: MEDIC‐PCR

Shrestha,

Kim,

Han

et al. 2023

Advanced Science

View full text Add to dashboard Cite

The COVID‐19 outbreak has caused public and global health crises. However, the lack of on‐site fast, reliable, sensitive, and low‐cost reverse transcription polymerase chain reaction (RT‐PCR) testing limits early detection, timely isolation, and epidemic prevention and control. Here, the authors report a rapid mobile efficient diagnostics of infectious diseases via on‐chip ‐RT‐quantitative PCR (RT‐qPCR): MEDIC‐PCR. First, the authors use a roll‐to‐roll printing process to accomplish low‐cost carbon‐black‐based disposable PCR chips that enable rapid LED‐induced photothermal PCR cycles. The MEDIC‐PCR can perform RT (3 min), and PCR (9 min) steps. Further, the cohort of 89 COVID‐19 and 103 non‐COVID‐19 patients testing is completed by the MEDIC‐PCR to show excellent diagnostic accuracy of 97%, sensitivity of 94%, and specificity of 98%. This MEDIC‐PCR can contribute to the preventive global health in the face of a future pandemic.

show abstract

Paper-based netlike rolling circle amplification (NRCA) for ultrasensitive and visual detection of SARS-CoV-2

Cited by 19 publications

References 31 publications

Precise Detection of Viral RNA by Programming Multiplex Rolling Circle Amplification and Strand Displacement

Precise Detection of Viral RNA by Programming Multiplex Rolling Circle Amplification and Strand Displacement

Recent Uses of Paper Microfluidics in Isothermal Nucleic Acid Amplification Tests

Mobile Efficient Diagnostics of Infectious Diseases via On‐Chip RT‐qPCR: MEDIC‐PCR

Contact Info

Product

Resources

About