1. An ultrasensitive and simple assay for the Hepatitis C virus using a reduced graphene oxide-assisted hybridization chain reaction.
- Author
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Fan J, Yuan L, Liu Q, Tong C, Wang W, Xiao F, Liu B, and Liu X
- Subjects
- Cell Line, Tumor, DNA chemistry, DNA genetics, DNA Probes chemistry, DNA Probes genetics, Fluoresceins chemistry, Fluorescence, Fluorescent Dyes chemistry, Graphite chemical synthesis, HEK293 Cells, Humans, Limit of Detection, Nucleic Acid Amplification Techniques methods, Nucleic Acid Hybridization, Oxidation-Reduction, Proof of Concept Study, RNA, Viral genetics, Spectrometry, Fluorescence methods, Biological Assay methods, Biosensing Techniques methods, Graphite chemistry, Hepacivirus isolation & purification, RNA, Viral analysis
- Abstract
Hepatitis C virus (HCV) is a major cause of chronic liver disease, which affects 2-3% of the world population. Until now, the early detection of HCV has been a great challenge, especially for those who live in developing countries. In this study, we developed a novel and ultrasensitive assay for the detection of HCV RNA based on the reduced graphene oxide nanosheets (rGONS) and hybridization chain reaction (HCR) amplification technique. This detection system contains a pair of single fluorophore-labeled hairpin probes that can freely exist in the solution in the absence of target RNA. The introduction of target RNA can robustly trigger a HCR with the two probes and produce long nanowires containing a double-stranded structure. The weak adsorption to rGONS makes the long nanowires emit a strong fluorescence. Using this enzyme-free amplification strategy, we developed a new method for the HCV RNA assay with a detection limit of 10 fM, which is far more sensitive than the common GO-based fluorescence method. Furthermore, the new method exhibits high selectivity for the discrimination of perfectly complementary and mismatched sequences. Finally, the new method was successfully used as a HCV RNA assay in biological samples with a strong anti-interference capability in complicated environments. In summary, these remarkable characteristics of the new method highlight its potential use in a clinical sample primary screening.
- Published
- 2019
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