Studies on the fluorescence fiber-optic DNA biosensor using p-hydroxyphenylimidazo[f]1,10-phenanthroline ferrum(III) as indicator.
A complex Fe(phen)(2).PHPIP.3ClO(4).2H(2)O, where phen=1,10-phenanthroline and PHPIP=p-hydroxyphenylimidazo[f]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 x 10(-7) to 4.88 x 10(-6) M and a detection limit of 1.08 x 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.
Niu
Shu-yan
SY
College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, People's Republic of China.
Wang
Shao-juan
SJ
Shi
Chao
C
Zhang
Shu-sheng
SS
D015374:Biosensing Techniques; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000378:metabolism*; D005290:Ferric Compounds / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D005336:Fiber Optic Technology; D005453:Fluorescence; D009693:Nucleic Acid Hybridization; D055100:Optical Fibers; D010618:Phenanthrolines / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
D004277:DNA, Single-Stranded; D005290:Ferric Compounds; D010618:Phenanthrolines; C528475:p-hydroxyphenylimidazo(f)1,10-phenanthroline ferrum(III)