Amplification of H1N1 and H3N2 by asymmetric RT-PCR for direct downstream applications

Straightforward point-of-care devices are essential for easier diagnostics of infectious diseases like influenza. Recent integrated microfluidic systems that use Reverse Transcriptase - Polymerase Chain Reaction (RT-PCR) for amplifying viral nucleic acids have sensing (detection) module that is not directly coupled to the amplification module. This is attributed to the fact that sensing modules require single stranded (ss) nucleic acid for hybridization, but the resultant amplicon of conventional amplification is double stranded (ds). This double stranded amplicon needs to be processed before being used in sensing module. We propose asymmetric RT-PCR of H1N1 and H3N2 viral nucleic acids as straightforward method to generate ssDNA facilitating its use directly for downstream applications such as hybridization with biosensors.