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| Content Provider | PubMed Central |
|---|---|
| Author | Negri, Pierre Dluhy, Richard A. |
| Abstract | We have developed a method for the detection of genetic markers associated with high pathogenicity in influenza. The assay consists of an array of 5′-thiolated ssDNA oligonucleotides immobilized on the surface of a Ag nanorod substrate that serve as capture probes for the detection of synthetic RNA sequences coding for a genetic mutation in the influenza PB1-F2 protein. Hybridization of the DNA probes to their complementary RNA sequences was detected using surface-enhanced Raman spectroscopy (SERS). Multivariate statistical analysis was used to differentiate the spectra of the complementary DNA probe-RNA target hybrids from those of the non-complementary DNA probes containing a single base pair polymorphism. Hierarchical cluster analysis (HCA) was able to distinguish with 100% accuracy the spectra of the complementary DNA probe – RNA target from the spectra of the immobilized DNA probes alone, or the DNA probes incubated with non-complementary RNA sequences. Linearity of response and limits of sensitivity of the SERS-based assays were determined using a partial least squares (PLS) regression model; detection limits computed by PLS was determined to be ~10 nM. The binding affinity of the DNA probes to their complementary RNA sequences was confirmed using enzyme-linked immunosorbent assay (ELISA); however, the detection limits observed using ELISA were approximately 10× higher (~100 nM) than those determined by PLS analysis of the SERS spectra. |
| Related Links | http://dx.doi.org/10.1039/c3an00774j |
| Starting Page | 4877 |
| File Format | |
| ISSN | 00032654 |
| e-ISSN | 13645528 |
| Journal | The Analyst |
| Issue Number | 17 |
| Volume Number | 138 |
| Language | English |
| Publisher Date | 2013-01-01 |
| Access Restriction | Open |
| Subject Keyword | Analytical Chemistry Spectroscopy Electrochemistry Biochemistry Environmental Chemistry Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Spectroscopy Environmental Chemistry Analytical Chemistry Biochemistry Electrochemistry |
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