Dual signal amplification for microRNA-21 detection based on duplex-specific nuclease and invertase.
Xitong HuangZhiming XuJi-Hua LiuBo-Yang YuJiangwei TianPublished in: RSC advances (2020)
MicroRNA-21 (miRNA-21) is a significant biomarker which is closely related to some kinds of diseases, such as cancer, cardiovascular disease and kidney disease. Therefore, the detection of miRNA-21 is of great importance and can provide essential information for disease diagnosis. In this study, we report a facile, sensitive assay for miRNA-21 detection using personal glucose meters (PGM). Biotinylated DNA strand linked invertase (Inv) is conjugated on the surface of streptavidin-coated magnetic beads (MBs) to form a MBs-DNA-Inv complex. Target miRNA-21 in the detection system is captured by the MBs-DNA-Inv probe through DNA/RNA hybridization. The duplex-specific nuclease (DSN) enzyme specifically cleaves the DNA to recycle the target miRNA and release invertase, thereby triggering the dual signal amplification and ensuring high sensitivity. Besides, we establish a linear relationship between PGM and different concentrations of miRNA-21 in the range of 10 to 200 pM. The limit of detection is 1.8 pM, which is more sensitive than some of the previous reports. In addition, the biosensor exhibits excellent sequence selectivity and single-base mutation can be discriminated. Moreover, the expression of miRNA-21 is confirmed in urine from mice by our method, which is in good accordance with the qRT-PCR result. Therefore, a dependable, low-cost strategy for the detection of miRNA has been established and it meets the latest analytical demands for miRNA determination that is suitable for the public.
Keyphrases
- label free
- nucleic acid
- circulating tumor
- real time pcr
- loop mediated isothermal amplification
- single molecule
- cardiovascular disease
- cell free
- low cost
- type diabetes
- air pollution
- quantum dots
- blood pressure
- healthcare
- squamous cell carcinoma
- particulate matter
- photodynamic therapy
- adipose tissue
- heavy metals
- insulin resistance
- poor prognosis
- social media
- skeletal muscle
- transcription factor
- weight loss
- metabolic syndrome
- risk assessment
- reduced graphene oxide
- high resolution