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Fast Optimization of a Multiplex Influenza Identification Panel Using a Thermal Gradient


The year 2009 was marked by the emergence of a novel influenza A (H1N1) virus that infects humans. There is a need to identify the different strains of influenza virus for purposes of monitoring the H1N1 strain pandemic and for other epidemiological and scientific purposes. In preparation for an accurate and rapid identification of H1N1, a laboratory in Singapore established a multiplex influenza identification panel that detects H1N1 among influenza A subtype H1 (flu A), subtype H3 (H3N2) and influenza B (flu B) viruses. The panel uses a 5' nuclease (TaqMan) probe–based four-target multiplex real-time PCR assay.

In this tech report we illustrate the use of the thermal gradient feature on the CFX96 real-timePCR detection system to optimize reverse transcription (RT) and fast PCR conditions for a four-target multiplex PCR assay.

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