New Method For Influenza Virus Detection Based On Quantum Dots

Influenza is a viral respiratory pathogen responsible for frequent seasonal epidemics. Recently, influenza viruses have been isolated from a variety of animals, including humans, pigs, horses, sea mammals, and birds. All avian influenza viruses are classified as type A, and belong to the Orthomyxoviridae family. Further subtyping is based on the antigenicity of two surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). HA and NA, which located on the surface of virus, play important role during the virus infection. Currently,16 HA (H1-H16) and 9 NA (N1-N9) subtypes have been identified among influenza A viruses. Recently, major human influenza viruses, such as H5N1 and H1N1, have globally circulated. Subtyping of the viruses is essential for diagnosis and effective antiviral treatment. Here we present a new detection protocol for the influenza virus using quantum dots (QDs)Firstly, QDs conjugated with oligonucleotide probes were used to produce fluorescent signals, whereas MBs that were also conjugated with the probes were used to isolate and concentrate the signals. The presence of target viral RNAs led to the sandwich hybridization between the functionalized QDs and MBs. One-step hybridization facilitated the simultaneous subtype determination and quantification of the influenza virus.We also prepared and purified polyclonal antibody correspond to HA, NA. Then the QDs and antibodies were coupled via the amido bond. Based on the antigen-antibody interaction, the purified antibodies couple with QDs recongnize the viral surface antigens. Meanwhile the fluorescent signals of QDs were detected to achieve the goal of detection and subtyping.Upon the superiorities of QDs, these detection systems may be also applicable for other DNA and RNA viruses and may function as a highly efficient clinical diagnosis kit for use in a doctor's office.