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Updated: Jul 18, 2026
High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
Evaluation of clinical specimens for influenza A virus positivity using various diagnostic methods
E Varecková1, H Blaskovicová, M Gocník
1Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 845 05 Bratislava, Slovak Republic. viruevar@savba.sk
Abstract:
The diagnostic method for Influenza A virus, utilizing the SERION ELISA Antigen kit (SERION EIA), if results were evaluated according to the manufacturer's instructions, has repeatedly failed to detect a great number of clinical samples positive by virus isolation and RT-PCR. Therefore we compared the SERION EIA with the one-step 44/107L-Px immunocapture enzyme immunoassay (44/107L-Px EIA), developed in our laboratory (Tkácová and Varecková, J. Virol. Methods 60, 65-71, 1996). Seventy-three clinical specimens, of which 65 were positive by virus isolation (used as reference method), were tested by both EIAs. By the SERION EIA, out of the 65 reference-positive samples only 8 (12%) were positive, 5 (8%) were ambiguous, and 52 (80%) were negative, which corresponded to the sensitivity of 12%. On the contrary, the sensitivity of the 44/107L-Px EIA was 74%. However, the calculation of cut-off values for the evaluation of positivity of clinical specimens in these two assays were not the same. If the evaluation procedure used for the 44/107L-Px EIA was applied to the SERION EIA, the sensitivity and the specificity of both EIAs became comparable, namely 71% and 100% for the SERION EIA and 74% and 100% for the 44/107L-Px EIA, respectively. From these results it follows that not the detection ability of the SERION EIA, but the evaluation procedure recommended by its manufacturer led to a loss of large number of positive specimens.
Insights
The SERION ELISA Antigen kit (SERION EIA) showed low sensitivity for Influenza A virus detection. Adjusting the evaluation method significantly improved its performance, matching a custom assay.
Area of Science:
- Virology
- Immunology
- Diagnostic Assay Development
Background:
- The SERION ELISA Antigen kit (SERION EIA) has demonstrated poor performance in detecting Influenza A virus in clinical samples.
- Virus isolation and RT-PCR confirmed positive results were missed by the SERION EIA when used per manufacturer's instructions.
Purpose of the Study:
- To compare the diagnostic performance of the SERION EIA with a laboratory-developed immunocapture enzyme immunoassay (44/107L-Px EIA).
- To evaluate the impact of different evaluation procedures on the sensitivity and specificity of both assays for Influenza A virus detection.
Main Methods:
- Seventy-three clinical specimens were tested using both the SERION EIA and the 44/107L-Px EIA.
- Virus isolation served as the reference method for determining true positive samples.
- Sensitivity and specificity were calculated for both assays under different evaluation criteria.
Main Results:
- The SERION EIA exhibited a low sensitivity of 12% when evaluated using the manufacturer's recommended cut-off values.
- The 44/107L-Px EIA demonstrated a sensitivity of 74%.
- When the evaluation procedure of the 44/107L-Px EIA was applied to the SERION EIA, its sensitivity increased to 71% with 100% specificity, comparable to the 44/107L-Px EIA's 74% sensitivity and 100% specificity.
Conclusions:
- The poor performance of the SERION EIA is attributed to its recommended evaluation procedure, not its inherent detection capability.
- Optimizing the cut-off value calculation significantly enhances the diagnostic accuracy of the SERION EIA for Influenza A virus.
- The study highlights the critical role of appropriate data interpretation in immunoassay diagnostics.
