An innovative gold nanorod (GNR) array biochip was developed to systematically investigate the localized surface plasmon resonance (LSPR)-coupled fluorescence enhancement for signal amplification in molecular beacon detection. An ordered GNR assembly in vertical standing array on a glass surface was fabricated as plasmonic substrates, resulting in dramatically intensified LSPR between adjacent nanoparticles as compared to that from an ensemble of random nanorods. We have shown that the plasmonic response of the nanoarray can be tuned by the proper choice of GNR size to overlap the fluorophore excitation and emission wavelengths greater than 600 nm. Plasmon-induced fluorescence enhancement was found to be distance-dependent with the competition between quenching and enhancement by the metal nanostructures. The augmented fluorescence enhancement by the GNR array can efficiently overcome the quenching effect of the gold nanoparticle even at close proximity. The enhancement correlates with the spectral overlap between the fluorophore excitation/emission and the plasmonic resonance of the GNR array, indicating a surface-plasmon-enhanced excitation and radiative mechanism for the amplification. From these results, the applicability of the ordered GNR array chip was extended to molecular fluorescence enhancement for practical use as a highly functional and ultrasensitive plasmonic DNA biochip in molecular beacon fashion.