Electrochemical genosensor for the rapid detection of GMO using loop-mediated isothermal amplification.
In this study, we are reporting for the first time an efficient, accurate and inexpensive rapid detection system which employs the integration of isothermal amplification and subsequent analysis of unpurified amplicons by an electrochemical system. In our experiments, loop-mediated isothermal amplification (LAMP) with its higher efficiency than PCR was performed at a constant temperature (65 degrees C). Amplification products were combined with a redox active molecule Hoechst 33258 [H33258, 2'-(4-hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bi(1H-benzimidazole)] and analyzed by a DNA stick (DS) which is integrated with a disposable electrochemical printed (DEP) chip using linear sweep voltammetry (LSV). The DNA minor groove binding of the H33258 molecule causes a significant drop in the peak current intensity of the H33258 oxidation. The phenomenon of DNA binding induced by H33258, in addition to changes in the anodic current peak, was used to detect maize CBH 351 variety (StarLink). Since laborious probe immobilization was not required, and amplification and detection were performed on a single device, our biosensor eliminates potential cross-contamination. We believe that this type of sensor will have an unprecedented impact for environmental protection.
Ahmed
Minhaz Uddin
MU
School of Materials Science, Japan Advanced Institute of Science and Technology (JAIST), 1-1 Asahidai, Nomi City, Ishikawa 923-1292, Japan.
Saito
Masato
M
Hossain
M Mosharraf
MM
Rao
S Ramachandara
SR
Furui
Satoshi
S
Hino
Akihiro
A
Takamura
Yuzuru
Y
Takagi
Masahiro
M
Tamiya
Eiichi
E
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D055664:Electrochemical Techniques / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation* / Q000379:methods; D030821:Plants, Genetically Modified / Q000235:genetics*; D003313:Zea mays / Q000235:genetics*