Overview

This practical guide details the essential methodologies for the isolation, identification, and quantification of Influenza A viruses, specifically focusing on the H1N1pdm09 strain using MDCK cell culture systems. It provides a comprehensive workflow that includes standardized protocols for the maintenance and subculturing of MDCK cells, viral propagation techniques, and molecular confirmation via RT-PCR targeting the HA gene. Furthermore, the text offers in-depth instruction on viral quantification methods, employing Hemagglutination (HA) assays to determine total viral titers and TCID50 assays using the Reed-Muench method for calculating infectious titers. By combining theoretical background with rigorous experimental data and step-by-step procedural flowcharts, this work serves as a vital resource for researchers and laboratory professionals establishing robust platforms for influenza virus characterization and antiviral drug discovery.

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