An Inexpensive and Accurate Reverse Transcription-PCR–Melting Temperature Analysis Assay for Real-Time Influenza Virus B Lineage Discrimination

Abstract

In this work, we describe a SYBR-Green one-step reverse transcription-PCR protocol coupled with a melting temperature analysis (RT-PCR- T m ), which allows the discrimination of influenza B lineages Yamagata and Victoria. The assay is performed using a regular real-time thermocycler and is based on differences in melting temperature ( T m ) of a 131-bp amplicon, obtained from a conserved region of hemagglutinin gene.