Modified-RNA aptamer-based sensor for competitive impedimetric assay of neomycin B

被引:95
作者
de-los-Santos-Alvarez, Noemi [1 ]
Lobo-Castanon, Maria Jesus [1 ]
Miranda-Ordieres, Arturo J. [1 ]
Tunon-Blanco, Paulino [1 ]
机构
[1] Univ Oviedo, Dept Quim Fis & Analit, Oviedo 33006, Spain
关键词
D O I
10.1021/ja0689482
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
The first label-free modified RNA-aptasensor for the detection of small molecules in biological samples is presented. A competitive displacement assay was applied to the detection of aminoglycoside neomycin B in whole milk using a fully 2'-O-methylated RNA aptamer with faradaic impedance spectroscopic (FIS) detection. Neomycin B in solution displaces the aptamer from its complex with the SAM-immobilized neomycin B. The reusable aptasensor is capable of discriminating neomycin B from paromomycin, which differs from it in the substitution of a single amine group with a hydroxyl one. We show that the modified endonuclease-resistant RNA aptamer maintains the exquisite selectivity of the natural aptamer and allows the examination of biological samples of high protein content. This detection scheme can be readily adapted to the detection of other aminoglycosides and small molecules.
引用
收藏
页码:3808 / +
页数:3
相关论文
共 23 条
[1]  
Baker BR, 2006, J AM CHEM SOC, V128, P3138, DOI 10.1021/ja056957p
[2]   Displacement enzyme linked aptamer assay [J].
Baldrich, E ;
Acero, JL ;
Reekmans, G ;
Laureyn, W ;
O'Sullivan, CK .
ANALYTICAL CHEMISTRY, 2005, 77 (15) :4774-4784
[3]   Natural and engineered nucleic acids as tools to explore biology [J].
Breaker, RR .
NATURE, 2004, 432 (7019) :838-845
[4]   Recognition of a cognate RNA aptamer by neomycin B: quantitative evaluation of hydrogen bonding and electrostatic interactions [J].
Cowan, JA ;
Ohyama, T ;
Wang, DQ ;
Natarajan, K .
NUCLEIC ACIDS RESEARCH, 2000, 28 (15) :2935-2942
[5]   INVITRO SELECTION OF RNA MOLECULES THAT BIND SPECIFIC LIGANDS [J].
ELLINGTON, AD ;
SZOSTAK, JW .
NATURE, 1990, 346 (6287) :818-822
[6]   Binding of neomycin-class aminoglycoside antibiotics to the A-site of 16 S rRNA [J].
Fourmy, D ;
Recht, MI ;
Puglisi, JD .
JOURNAL OF MOLECULAR BIOLOGY, 1998, 277 (02) :347-362
[7]   RNA as a drug target: chemical, modelling, and evolutionary tools [J].
Hermann, T ;
Westhof, E .
CURRENT OPINION IN BIOTECHNOLOGY, 1998, 9 (01) :66-73
[8]   Development of ELISA and immunochromatographic assay for the detection of neomycin [J].
Jin, Y ;
Jang, JW ;
Lee, MH ;
Han, CH .
CLINICA CHIMICA ACTA, 2006, 364 (1-2) :260-266
[9]   Automated microarray system for the simultaneous detection of antibiotics in milk [J].
Knecht, BG ;
Strasser, A ;
Dietrich, R ;
Märtlbauer, E ;
Niessner, R ;
Weller, MG .
ANALYTICAL CHEMISTRY, 2004, 76 (03) :646-654
[10]   Trace analysis of aminoglycoside antibiotics in bovine milk by MEKC with LIF detection [J].
Manuel Serrano, Juan ;
Silva, Manuel .
ELECTROPHORESIS, 2006, 27 (23) :4703-4710