A Biochemiluminescent Assay for Rapid Diagnosis of Influenza

Abstract Background: A biochemiluminescent assay of influenza diagnosis is presented. The assay diagnoses influenza based on detection of the influenza viral neuraminidase activity. An instrument designed for the assay is also reported. This assay solves the problem that current influenza virus diagnosis assays are susceptible to virus mutation. Methods: A luciferase-based complex is synthesized as biochemiluminescent substrate. The substrate is cleaved to free luciferin with presence of influenza neuraminidase in specimen. Luciferase is oxidized to oxyluciferin with luciferin as catalyzer resulting in luminescence, which is proportional to the neuraminidase activity and measured by instrument. The instrument uses a photomultiplier tube (PMT) as sensor, with 24 test channels. Fine optical arrangements enable the instrument with high sensitive and accuracy. Results: A total of 389 clinical specimens were collected to evaluate the performance of the assay in clinical settings. This assay had a sensitivity and specificity of 95.92% (95% confidence interval: 91.38%-98.12%) and 97.93% (95% confidence interval: 95.26%-99.11%), respectively, compared to the colloidal gold assay. Conclusion: As a biochemiluminscence assay, this assay is advantageous in sensitivity and specificify. It does not require any washing and separation steps, which makes the instrument simple in design and easy to operate or maintenance. The assay is suitable for the rapid diagnosis of influenza virus in POC settings.