Abstract
We report application of lanthanide nanoparticles for DNA quantification in a microarray platform as a substitute for conventional organic fluorophores. A non-PCR based DNA microarray assay for quantifying bacteria capable of biodegrading methyl tertiary-butyl ether (MTBE) was demonstrated. Probe DNA was immobilized on a glass surface, hybridized with biotinylated target DNA and subsequently incubated with Neutravidin-biofunctionalized nanoparticles. The fluorescence spot intensities, measured by a commercial laser scanner, show a linear relationship (R 2 = 0.98) with bacterial 16S rDNA over a range of target DNA concentrations, while the background fluo-rescence remained low. In addition, nanoparticles fluorescence shows a stronger intensity than Quasar570 (Cy3). Present sensitivity of the assay is 10 pM of target DNA. The selectivity of the DNA-nanoparticle-probes to discriminate a non-target DNA with two base pairs mismatch in the 16S rDNA gene sequence was shown. The use of Eu:Gd 2O 3 nanoparticles as biolabels provides a relatively non-toxic, inexpensive, rapid and sensitive alternative to the materials currently used in DNA microarrays.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2462-2467 |
| Number of pages | 6 |
| Journal | Journal of Nanoscience and Nanotechnology |
| Volume | 8 |
| Issue number | 5 |
| State | Published - May 2008 |
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Keywords
- DNA microarrays
- Lanthanide europium oxide
- Methylibium petroleiphilum PM1
- MTBE
- Nanoparticles
ASJC Scopus subject areas
- Chemistry(all)
- Materials Science(all)
- Materials Science (miscellaneous)
- Engineering (miscellaneous)
Cite this
Luminescent lanthanide nanoparticles as labels in DNA microarrays for quantification of methyl tertiary butyl ether degrading bacteria. / Son, Ahjeong; Nichkova, Mikaela; Dosev, Dosi; Kennedy, Ian M.; Hristova, Krassimira R.
In: Journal of Nanoscience and Nanotechnology, Vol. 8, No. 5, 05.2008, p. 2462-2467.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Luminescent lanthanide nanoparticles as labels in DNA microarrays for quantification of methyl tertiary butyl ether degrading bacteria
AU - Son, Ahjeong
AU - Nichkova, Mikaela
AU - Dosev, Dosi
AU - Kennedy, Ian M.
AU - Hristova, Krassimira R.
PY - 2008/5
Y1 - 2008/5
N2 - We report application of lanthanide nanoparticles for DNA quantification in a microarray platform as a substitute for conventional organic fluorophores. A non-PCR based DNA microarray assay for quantifying bacteria capable of biodegrading methyl tertiary-butyl ether (MTBE) was demonstrated. Probe DNA was immobilized on a glass surface, hybridized with biotinylated target DNA and subsequently incubated with Neutravidin-biofunctionalized nanoparticles. The fluorescence spot intensities, measured by a commercial laser scanner, show a linear relationship (R 2 = 0.98) with bacterial 16S rDNA over a range of target DNA concentrations, while the background fluo-rescence remained low. In addition, nanoparticles fluorescence shows a stronger intensity than Quasar570 (Cy3). Present sensitivity of the assay is 10 pM of target DNA. The selectivity of the DNA-nanoparticle-probes to discriminate a non-target DNA with two base pairs mismatch in the 16S rDNA gene sequence was shown. The use of Eu:Gd 2O 3 nanoparticles as biolabels provides a relatively non-toxic, inexpensive, rapid and sensitive alternative to the materials currently used in DNA microarrays.
AB - We report application of lanthanide nanoparticles for DNA quantification in a microarray platform as a substitute for conventional organic fluorophores. A non-PCR based DNA microarray assay for quantifying bacteria capable of biodegrading methyl tertiary-butyl ether (MTBE) was demonstrated. Probe DNA was immobilized on a glass surface, hybridized with biotinylated target DNA and subsequently incubated with Neutravidin-biofunctionalized nanoparticles. The fluorescence spot intensities, measured by a commercial laser scanner, show a linear relationship (R 2 = 0.98) with bacterial 16S rDNA over a range of target DNA concentrations, while the background fluo-rescence remained low. In addition, nanoparticles fluorescence shows a stronger intensity than Quasar570 (Cy3). Present sensitivity of the assay is 10 pM of target DNA. The selectivity of the DNA-nanoparticle-probes to discriminate a non-target DNA with two base pairs mismatch in the 16S rDNA gene sequence was shown. The use of Eu:Gd 2O 3 nanoparticles as biolabels provides a relatively non-toxic, inexpensive, rapid and sensitive alternative to the materials currently used in DNA microarrays.
KW - DNA microarrays
KW - Lanthanide europium oxide
KW - Methylibium petroleiphilum PM1
KW - MTBE
KW - Nanoparticles
UR - http://www.scopus.com/inward/record.url?scp=84857899407&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84857899407&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:84857899407
VL - 8
SP - 2462
EP - 2467
JO - Journal of Nanoscience and Nanotechnology
JF - Journal of Nanoscience and Nanotechnology
SN - 1533-4880
IS - 5
ER -