Time-resolved fluorescence energy transfer (TR-FRET) assay for detecting exposure to an infectious organism

摘要

A method for detecting the presence of a diagnostic moiety indicative of exposure to a Bovine Viral Diarrhoea virus infectious organism in a biological sample taken from a human or animal, said method comprising;
(a) adding to said sample a first fluorescently labelled reagent which binds said diagnostic moiety, and a second fluorescently labelled reagent which either binds said diagnostic moiety in addition to said first fluorescently labelled reagent, or which binds the first fluorescently labelled reagent or a complex comprising the first fluorescently labelled reagent in competition to the said diagnostic moiety, wherein a label on one of the first or second fluorescently labelled reagent acts as a fluorescent energy donor compound and wherein the other of the first or second fluorescently labelled reagent acts as a fluorescent energy acceptor compound which is able to accept fluorescent energy from said donor compound;
(b) exciting the fluorescent energy donor compound by illuminating with light of a wavelength which is absorbed by said fluorescent energy donor compound;
(c) measuring fluorescent signal emitted by said fluorescent energy acceptor compound as a result of its absorption of the fluorescent energy from the donor compound after a time delay; and
(d) relating the results to the presence or absence of diagnostic moiety in said sample.