Studies on the Fluorescence Fiber-Optic DNA Biosensor Using <Emphasis Type="Italic">p</Emphasis>-Hydroxyphenylimidazo[f]1,10-phenanthroline Ferrum(III) as Indicator |
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Authors: | Shu-yan Niu Shao-juan Wang Chao Shi Shu-sheng Zhang |
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Institution: | (1) College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, 266042, People’s Republic of China |
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Abstract: | A complex Fe(phen)2·PHPIP·3ClO4·2H2O, where phen = 1,10-phenanthroline and PHPIP = p-hydroxyphenylimidazof]1,10-phenanthroline, was synthesized and acted as a good fluorescence indicator based on its interaction
with double-duplex DNA. Then a fiber-optic DNA biosensor of fluorimetric detection was developed based on the recognition
of target DNA in DNA hybridization assays. A probe ssDNA was covalently immobilized onto the surface of quartz optical fibers
and then the probe ssDNA hybridized with complementary ssDNA introduced into the local environment of the sensor. The hybridization
with complementary strands was monitored in real time by fluorimetric detection. Several factors affecting the probe immobilization,
target DNA hybridization, and indicator binding reactions were optimized to maximize the sensitivity and shorten the assay
time. Using this method, a sequence of the 16-mer oligonucleotides could be quantified over the range from 4.98 × 10−7 to 4.88 × 10−6 M and a detection limit of 1.08 × 10−7 M. And the designed optic-fiber biosensor could be conveniently regenerated by thermal denature. The utility of the novel
hybridization indicator could provide a simple, rapid, low toxicity and reusable detection. |
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Keywords: | Fe(phen)2·PHPIP·3ClO4·2H2O Fiber-optic biosensor Fluorimetric detection Oligonucleotides DNA hybridization |
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