Highly sensitive and portable mRNA detection platform for early cancer detection
| Autor: | Wenqiong Su, Laikuan Zhu, Kan Wang, Xiao Zhi, Jie He, Xianting Ding, Hongxia Li, Antony R. Warden, Guangxia Shen |
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| Rok vydání: | 2021 |
| Předmět: |
mRNA
Biomedical Engineering Metal Nanoparticles Pharmaceutical Science Medicine (miscellaneous) Bioengineering Lateral flow assay Applied Microbiology and Biotechnology Extracellular Vesicles chemistry.chemical_compound Glypicans Pancreatic cancer Medical technology Biomarkers Tumor medicine Humans RNA Messenger R855-855.5 Early Detection of Cancer Detection limit Messenger RNA Research COVID-19 medicine.disease Molecular medicine Biomarker (cell) Cell biology Pancreatic Neoplasms Point-of-care testing chemistry Colloidal gold Duplex (building) Early-stage cancer detection Molecular Medicine Gold TP248.13-248.65 Glypican-1 DNA Biotechnology |
| Zdroj: | Journal of Nanobiotechnology Journal of Nanobiotechnology, Vol 19, Iss 1, Pp 1-10 (2021) |
| ISSN: | 1477-3155 |
| DOI: | 10.1186/s12951-021-01039-4 |
| Popis: | Pancreatic 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.). Supplementary Information The online version contains supplementary material available at 10.1186/s12951-021-01039-4. |
| Databáze: | OpenAIRE |
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