| Summary: | 碩士 === 東海大學 === 化學系 === 103 === The abuse of antibiotics increase pathogens to produce resistance gene then the multidrug resistance gene (MDR gene) emerged with the situation become worse. To prevent the MDR gene spreading, it have to make the right decision for taking the antibiotics. For this reason, we take the superbug having New delhi metallo-beta-lactamase (NDM) be the sample to develop an electrobiosensor which modified by electrochemical grafting diazonium compounds. Construct a rapid-modifying and precise platform to detect the blaNDM.
The accurate analysis was supported by detect two encoding gene from binding site of NDM and one specific gene of blaNDM, when simultaneously sensing these three genetic sequences to generate output current. On the other side, we synthesize two aniline compounds, 3-((4-aminophenyl)dimethylammonio)propane-1-sulfonate (SC) and 4-amino-N-(2-(4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)benzamido)ethyl)benzamide (MD), to use the MD coupling the thiol-modified nucleotide probe, and SC to resist non-specific binding. Diazotized MD and SC to product both aryl diazonium compounds then using electrochemical grafting method to deposition two molecules on the gold electrode. Before to construct the MD and SC modified platform, we use the commercial compounds, para-phenylenediamine (PPDA) coupling m-maleimidobenzoyl-N-hydoxysuccinimide ester (MBS), and preparing the anti-fouling modified by self-assembly monolayer (SAM) system, then to analyze the electrochemical character of ary diazonium-modified electrode and the effect of modified concentration and cycles with nucleotide sequences detection. After that, consider the conditions of modified PPDA to deposite the MD and SC platform, then study the ways of modified for two compounds (two step or co-deposition) and the ratio of concentration with two compounds to compare the sensing.
Finally, this platform modified by co-deposition MD and SC with the ratio for 1:1 in total concentration 0.1 mM, then obtain the sensing for S/N ratio with 19. This way is better than two step modified process with higher S/N ratio and reproducible. In our research, we succeed using electrochemical grafting method to functionalized surface by diazonium compounds. Comparing to the traditional method of functionalized surface, self-assembly monolayer , the platform have short-term for modified, simple modifying-steps and few chemical material of using, and offer a nice or better electrobiosensor for detecting the MDR gene.
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