A label-free method for the detection of specific DNA sequences using gold nanoparticles bifunctionalized with a chemiluminescent reagent and a catalyst as signal reporters

Abstract

Sensitive, specific, simple, fast, and low-cost DNA detection methods are extremely important in clinical diagnostics, gene therapy, and a variety of biomedical studies. In this work, we developed a general method for the detection of specific DNA sequences from Mycobacterium tuberculosis (TB), hepatitis B virus (HBV), and myelocytomatosis viral oncogene (v-myc) using gold nanoparticles bifunctionalized with both a chemiluminescent (CL) reagent and a catalytic metal complex as signal reporters and a DNA strand complementary to the target as the capture probe. In this CL method, a biotinylated single-strand DNA capture probe was immobilized in a streptavidin-coated microwell. Upon the addition of the target single-strand DNA, the capture probe hybridized with the target DNA. After adding the bifunctionalized gold nanoparticles and H₂O₂, a well-defined CL signal was obtained, and the CL intensity was observed to change as the target DNA concentration was increased. It was possible to determine the concentration of the target TB single-strand DNA in the range 1.0 × 10^-13-1.0 × 10^-8 M with a detection limit of 4.8 × 10^-14 M. HBV single-strand DNA and v-myc single-strand DNA could also be determined in the range 1.0 × 10^-11-1.0 × 10^-8 M with detection limits of 5.9 × 10^-12 M and 8.0 × 10^-12 M, respectively, using this CL technique. The method reported in this paper is the first label-free CL method for the determination of specific DNA sequences to utilize gold nanoparticles bifunctionalized with both a CL reagent and a catalytic metal complex. The sensitivity of this CL method is superior to those of most previously reported label-free methods. Compared with methods that use polymerase chain reaction amplification, this label-free CL method is much simpler, faster, and more economic. This work has thus demonstrated a simple and fast scanning strategy for the detection of specific DNA sequences related to diseases. Graphical Abstract Schematic illustration of label-free CL method for detection of specific DNA sequences.

Keywords: Bifunctionalized gold nanoparticles; Chemiluminescence; DNA; Determination; Metal complexes; N-(Aminobutyl)-N-(ethylisoluminol).