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Nucleic Acids Research, 1986, Vol. 14, No. 2 1017-1028
© 1986

Articles

Time-resolved fluorometry: a sensitive method to quantify DNA-hybrids

Ann-Christine Syvänen, Paul Tchen1, Marjut Ranki and Hans Söderlund2

Orion Genetic Enginerring Laboratory Valimotie 7, 00380 Helsinki, Finland 1Institut National de la Santé et de la Recherche Medicale U. 155, Château de Longchamp, 75016 Paris, France 2Recombinant DNA Laboratory, University of Helsinki Valimotie 7, 00380 Helsinki, Finland

Received October 16, 1985. Revised December 3, 1985. Accepted December 3, 1985.

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.


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