Systems and methods for enhancing fluorescent detection of target molecules in a test sample are for use with an irradiating device. First fluorophores are provided for absorption of EMF radiation, and emission of a first signal. Second fluorophores are provided for partial absorption of the first signal, and emission of a second signal distinguishable from the first signal. The fluorophores are combined with the test sample, and secured to the target molecules and relative to one another. After the first fluorophores receive the EMF radiation from the irradiating device, the first signal is detected, together with the second spectral signal if the target molecules are present in the test sample.