Quantum-dots-based fluoroimmunoassay for the rapid and sensitive detection of avian influenza virus subtype H5N1

摘要

The continuous spread of highly pathogenic avian influenza virus (AIV) subtype H5N1 is threatening the poultry industry and human health worldwide. Rapid and sensitive diagnostic methods are required for the H5N1 surveillance. In this study, the fluorescent (FL) probe of CdTe quantum dots (QDs) was designed using covalently linked rabbit anti-AIV H5N1 antibody. Based on these QD-antibody conjugates, a novel sandwich FL-linked immunosorbent assay (sFLISA) was developed for H5N1 viral antigen detection. The sFLISA allowed for H5N1 viral antigen determination in a linear range of 8.0 × 10~(-3) to 5.1 × 10~(-1) μmL~(-1) with the limit of detection (LOD) of 1.5 × 10~(-4)μmL~(-1). In comparison with virus isolation for 103 clinic samples, the sensitivity and specificity of sFLISA were found to be 93.6 and 91.1% respectively. The sFLISA supplied a novel approach to rapid and sensitive detection of AIV subtype H5N1 and showed great potential for biological applications in immunoassays.