title: Two-stage cyclic enzymatic amplification method for ultrasensitive electrochemical assay of microRNA-21 in the blood serum of gastric cancer patients.
abstract: MicroRNA (miRNA) is riveting nowadays due to its close relevance to human malignancies. Exploiting a fast and convenient biosensor for sensitive and specific detecting miRNA is necessary and it has significant meaning for oncology. In this work, to understand the relationship between miRNA-21 and gastric cancer, we employ T4 RNA ligase 2 to initiate specific ligation reaction depending on the sequence of target RNA, and T7 exonuclease to catalyze the first stage cyclic enzymatic amplification method (CEAM) specifically, which also resting on the target RNA sequence, and the second stage CEAM to further amplify the response resulting from the initial target RNA. Our two-stage CEAM strategy can detect miRNA-21 as low as 0.36fM with remarkable specificity, and most importantly, it can be used to inspect the expression level of miRNA-21 in blood serum of gastric cancer patients. This will offer new perspective for figuring out the pathways of miRNA-21 involving in cancer.
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian, Shandong 271018, PR China.
authors.forename: Bingchen
authors.initials: B
authors.lastname: Li
authors.affiliation: The Tumor Center of Taian City Central Hospital, Taian, Shandong 271000, PR China.
authors.forename: Fei
authors.initials: F
authors.lastname: Liu
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian, Shandong 271018, PR China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Peng
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian, Shandong 271018, PR China.
authors.forename: Yunlei
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian, Shandong 271018, PR China.
authors.forename: Wenxuan
authors.initials: W
authors.lastname: Fan
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian, Shandong 271018, PR China; State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, PR China. Electronic address: yinhs@sdau.edu.cn.
authors.forename: Huanshun
authors.initials: H
authors.lastname: Yin
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian, Shandong 271018, PR China. Electronic address: ashy@sdau.edu.cn.
authors.forename: Shiyun
authors.initials: S
authors.lastname: Ai
authors.affiliation: State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, PR China. Electronic address: zhangxs@sdau.edu.cn.
authors.forename: Xiansheng
authors.initials: X
authors.lastname: Zhang
mesh_terms: D015374:Biosensing Techniques; D055664:Electrochemical Techniques / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D035683:MicroRNAs / Q000097:blood*; D013274:Stomach Neoplasms / Q000097:blood*
chemical_list: C524940:MIRN21 microRNA, human; D035683:MicroRNAs
keywords: Electrochemical biosensor; Gastric cancer; MicroRNA-21; T4 RNA ligase 2; T7 exonuclease