A highly sensitive immunosensor with 1,1,’-oxalyldiimidazole chemiluminescence (ODI-CL) detection was developed for the rapid monitoring of influenza A viruses such as H1N1, H3N2, and H5N1 in a sample. A specific antibody, capable of capturing an influenza A virus with good selectivity, was immobilized on the surface of polystyrene strip-well. A standard or sample (100 ml) added in the strip-well was incubated for 30 min at 37 °C. After the incubation, the strip-well was washed 3 times using PBST. Then, 4-Methylumbelliferyl-N-acetyl-α-D-neuraminic acid (MUNANA, 100 ml), known as fluorogenic substrate of neuraminidase, was added and incubated in the strip-well for a certain time (e.g., 3, 5, 10 min) to produce 4-Methylumbelliferone (4-MU) from the reaction between MUNANA and neuraminidase of influenza A virus. After the incubation, the solution (10 ml) containing 4-MU and MUNANA was transferred into a borosilicate test tube. The test tube was inserted into a detection cell of luminometer. Finally, the concentration of influenza A virus in a sample was quantified with relative CL intensity measured for 2 sec when ODI-CL reagents (e.g., H2O2, ODI) were consecutively added in the test tube. The relative CL was proportionally enhanced with the increase of influenza A virus concentration. The correlation between the new immunosensor and conventional enzyme immunoassay was studied. Additionally, the accuracy, precision, recovery, and reproducibility of the new immunosensor were studied to confirm whether it can be applied as an alternative analytical tool to replace time-consuming and expensive conventional methods.

Development of cost-effective chemiluminescent immunosensor for the rapid monitoring of influenza A viruses

254th National Meeting (2017)

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