Electrochemical competitive immunodetection of messenger RNA modified with N6-methyladenosine by using DNA-modified mesoporous PtCo nanospheres.

Autor: Ou X; Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People's Republic of China., Pu Q; Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People's Republic of China., Sheng S; Clinical Laboratory of Hospital Affiliated to Chengdu University, Chengdu, 610081, People's Republic of China., Dai T; Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People's Republic of China., Gou D; Clinical Laboratories, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People's Republic of China., Yu W; Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People's Republic of China., Yang T; Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People's Republic of China., Dai L; Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People's Republic of China., Yang Y; Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People's Republic of China. yangyujun@cqmu.edu.cn., Xie G; Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People's Republic of China. guomingxie@cqmu.edu.cn.
Jazyk: angličtina
Zdroj: Mikrochimica acta [Mikrochim Acta] 2019 Dec 09; Vol. 187 (1), pp. 31. Date of Electronic Publication: 2019 Dec 09.
DOI: 10.1007/s00604-019-4010-8
Abstrakt: N 6 -Methyladenosine (m 6 A) is the most abundant RNA modification in eukaryotic messenger RNA (mRNA). A highly sensitive electrochemical immunosensor is described for the determination of m 6 A-RNA. The method is based on the use of antibody (anti-m 6 A) and PtCo mesoporous nanospheres (MPNs). The analogously modified probe of type m 6 A-DNA-PtCo competes with m 6 A-RNA for antibodies on the gold electrode as an electrical signal probe. The electrical signal, best acquired at a working potential of -0.37 V (vs. Ag/AgCl) reflects the concentration of m 6 A. The PtCo MPNs catalyze the reduction of H 2 O 2 , and this amplifies the current and enhances sensitivity. The detection time of the assay is <1.5 h. Under optimal conditions, response is linear in the 0.005 to 100 nM m 6 A RNA concentration range, and the detection limit is 2.1 pM. The results obtained by this immunoassay with human cell lines are comparable to those obtained with a commercial kit. Graphical abstractSchematic representation of a method for electrochemical determination of m 6 A-modified mRNA. Anti-m 6 A Ab: antibody against m 6 A; BSA: bovine serum albumin; PtCo: PtCo mesoporous nanospheres; SH-m 6 A-DNA: DNA modified with both m 6 A and thiol groups; DPV: differential pulse voltammetry.
Databáze: MEDLINE
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