Luminescent lanthanide nanoparticles as labels in DNA microarrays for quantification of methyl tertiary butyl ether degrading bacteria.
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 (R2 = 0.98) with bacterial 16S rDNA over a range of target DNA concentrations, while the background fluorescence 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:Gd2O3 nanoparticles as biolabels provides a relatively non-toxic, inexpensive, rapid and sensitive alternative to the materials currently used in DNA microarrays.
Department of Land, Air, and Water Resources, University of California Davis, One Shields Avenue, Davis, California 95616, USA.
Ahjeong
A
Son
Mikaela
M
Nichkova
Dosi
D
Dosev
Ian M
IM
Kennedy
Krassimira R
KR
Hristova
D001419:Bacteria / Q000235:genetics / Q000378:metabolism*; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D053768:Metal Nanoparticles; D008738:Methyl Ethers / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D020411:Oligonucleotide Array Sequence Analysis; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics
D028581:Lanthanoid Series Elements; D008738:Methyl Ethers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; C043243:methyl tert-butyl ether