Optically encoded multifunctional nanospheres for one-pot separation and detection of multiplex DNA sequences

In this study, we report a simple method for simultaneous detection of multiplex DNA sequences, including complementary DNA (cDNA) sequences of HIV and HCV, DNA sequence of HBV, with QDs-encoded fluorescent nanospheres and nano-γ-Fe2O3-coated magnetic nanospheres. Detection was achieved on a fluorescence spectrophotometer without additional auxiliary instruments, and the detection limit was about 100 pM. Here, QDs-encoded fluorescent nanospheres (FNS) with different photoluminescent properties, and magnetic nanospheres (MNS) were separately fabricated by stepwise assembly of hydrophobic QDs or nano-γ-Fe2O3 on the surface of branched poly(ethylene imine) (PEI)-coated nanospheres in precisely controlled amounts, finally followed by silica encapsulation. FNS-labeled probe DNAs and MNS-labeled capture DNAs were used to hybridize with the corresponding targets at the same time. After magnetic separation, the sandwich-structured adducts were measured by fluorescence spectrophotometry. The results indicated that the targets could be detected with high sensitivity. This method is convenient, fast enough, and capable of high anti-interference. Therefore, it is expected to be used for simultaneous detection and separation of multiple targets at high levels of purity and throughput.