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.
Son
Ahjeong
A
Department of Land, Air, and Water Resources, University of California Davis, One Shields Avenue, Davis, California 95616, USA.
Nichkova
Mikaela
M
Dosev
Dosi
D
Kennedy
Ian M
IM
Hristova
Krassimira R
KR
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