mrna detection
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2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Hongxia Li ◽  
Antony R. Warden ◽  
Wenqiong Su ◽  
Jie He ◽  
Xiao Zhi ◽  
...  

AbstractPancreatic cancer, at unresectable advanced stages, presents poor prognoses, which could be prevented by early pancreatic cancer diagnosis methods. Recently, a promising early-stage pancreatic cancer biomarker, extracellular vesicles (EVs) related glypican-1 (GPC1) mRNA, is found to overexpress in pancreatic cancer cells. Current mRNA detection methods usually require expensive machinery, strict preservation environments, and time-consuming processes to guarantee detection sensitivity, specificity, and stability. Herein, we propose a novel two-step amplification method (CHAGE) via the target triggered Catalytic Hairpin Assembly strategy combined with Gold-Enhanced point-of-care-testing (POCT) technology for sensitive visual detection of pancreatic cancer biomarker. First, utilizing the catalyzed hairpin DNA circuit, low expression of the GPC1 mRNA was changed into amplification product 1 (AP1, a DNA duplex) as the next detection targets of the paper strips. Second, the AP1 was loaded onto a lateral flow assay and captured with the gold signal nanoparticles to visualize results. Finally, the detected results can be further enhanced by depositing gold to re-enlarge the sizes of gold nanoparticles in detection zones. As a result, the CHAGE methodology lowers the detection limit of mRNA to 100 fM and provides results within 2 h at 37 °C. Furthermore, we demonstrate the successful application in discriminating pancreatic cancer cells by analyzing EVs’ GPC1 mRNA expression levels. Hence, the CHAGE methodology proposed here provides a rapid and convenient POCT platform for sensitive detection of mRNAs through unique probes designs (COVID, HPV, etc.).


2021 ◽  
Vol 2 (3) ◽  
pp. 100711
Author(s):  
Giovanni H. Diaz ◽  
Stefan Heller
Keyword(s):  

2021 ◽  
pp. 113507
Author(s):  
Dongdong Liu ◽  
Wenhua Li ◽  
Mingzhu Yang ◽  
Lizhen Qiu ◽  
Hongru Pian ◽  
...  

2020 ◽  
Vol 167 ◽  
pp. 112474 ◽  
Author(s):  
Jeong Moon ◽  
Jaewoo Lim ◽  
Seoyoung Lee ◽  
Hye Young Son ◽  
Hyun Wook Rho ◽  
...  

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