| Summary: | 碩士 === 國立中正大學 === 化學所 === 97 === Using Fiber-Optic Localized Plasmon Resonance (FO-LPR) method,
we have developed a novel label-free biosensor for DNA and message
RNA (mRNA). Upon functionalization of the colloidal gold surface with
an oligonucleotide probe, the hybridization of the probe with
complementary DNA or mRNA was interrogated by the unique optical
properties of colloidal gold. The thiolated DNA was immobilized onto the
gold surface via the well-known self-assembly chemistry, where thiolated
DNA is assembled onto a gold surface followed by adsorption of a diluent
of an alkanethiol. The diluent not only helps to project the probe DNA off
the surface but also resists nonspecific adsorption onto the gold
nanoparticle surface.
Since the optical properties and hence the attenuated total reflection
spectrum of self-assembled gold colloids on the optical fiber changes
with the adsorbate induced refractive index of the environment near the
colloidal gold surface, the realization of the sensing platform is through
the measurement of intensity from the colloidal gold-modified optical
fiber. The limit of detection for DNA hybridization was
about 10-8~10-11M, depending on the number of base in the target DNA.
The FO-LPR method was further applied to detection of mRNA in
serum samples from patients. This study, we used real-time PCR to
quantify mRNA during HLA-B27 gene expression in patients with
Ankylosing Spondylitis and compared with results from FO-LPR method.
Results show that mRNA formed during HLA-B27 gene expression can
be detected and the sensor response correlate with results from real-time
PCR.
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