Time-resolved fluorometry: a sensitive method to quantify DNA-hybrids
- 26 January 1986
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 14 (2) , 1017-1028
- https://doi.org/10.1093/nar/14.2.1017
Abstract
Europium and other lanthanides can be excitated with UV-radiation, whereafter the energy is released as fluorescence, delayed in time up to 1 ms after the excitation. Eu can be used as a sensitive label in biological assays. Here we report on the application of time-resolved fluorometry to detect nucleic acid hybrids. The probe DNA was tagged with a hapten, either a fluorene or a sulfone group. After hybridization the probe DNA was detected by a two-step immunological assay with the second antibody labelled with Eu. The method is quantitative with a detection limit of 0.3 pg of actual target regions of immobilized adenovirus genomic DNA. The label was also used in sandwich hybridization, which allowed analyzing nasopharyngeal mucus for the presence of adenovirus.Keywords
This publication has 17 references indexed in Scilit:
- Sandwich hybridization as a convenient method for the detection of nucleic acids in crude samplesPublished by Elsevier ,2003
- Chemically modified nucleic acids as immunodetectable probes in hybridization experiments.Proceedings of the National Academy of Sciences, 1984
- Europium as a label in time-resolved immunofluorometric assaysAnalytical Biochemistry, 1984
- Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots.Proceedings of the National Academy of Sciences, 1983
- Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probesVirology, 1983
- NOVEL TEST FOR RAPID VIRAL DIAGNOSIS: DETECTION OF ADENOVIRUS IN NASOPHARYNGEAL MUCUS ASPIRATES BY MEANS OF NUCLEIC-ACID SANDWICH HYBRIDISATIONThe Lancet, 1983
- The making of strand-specific M13 probesGene, 1982
- Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes.Proceedings of the National Academy of Sciences, 1981
- Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase IJournal of Molecular Biology, 1977
- Protein a from Staphylococcus aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulinsFEBS Letters, 1972