Immobilization of gold nanoparticles with rhodamine to enhance the fluorescence resonance energy transfer between quantum dots and rhodamine; new method for downstream sensing of infectious bursal disease virus

摘要

Infectious bursal disease virus is a causative agent of one of the most important disease which causes frequent tragic disaster in the poultry industry all over the world. Therefore, in the present study a new fluorescence resonance energy transfer-based technique was developed to detect VP2 gene of infectious bursal disease virus using two oligonucleotide probes labeled with quantum dots and rhodamine- immobilized gold nanoparticles (AuNPs-Rh). Quantum dots labeled with an amino-modified first oligonucleotide, and AuNP5-Rh labeled with thiol-modified second oligonudeotides were added to the DNA targets upon which hybridization occurred. In the presence of target the AuNPs-Rh will be located in the vicinity of the quantum dots and leads to the fluorescence resonance energy transfer to be occurred and subsequently the fluorescence intensity of quantum dots was stimulated. The immobilization of rhodamine to the surface of AuNPs increased the fluorescence intensity of rhodamine. The maximum fluorescence resonance energy transfer efficiency for the developed sensor is monitored at a quantum dots-P-A/AuNPs-Rh-P-T molar ratio of 1:10. Moreover, the feasibility of the developed nanobiosensor was demonstrated by the detection of a synthetic 49-mer nucleotide derived from infectious bursal disease virus and the limit of detection was estimated as 3 x 10(-8) M. The developed DNA detection scheme is a simple, rapid and efficient technique which does not need excessive washing and separation steps. (C) 2018 Published by Elsevier B.V.