Nanomolecular detection of human influenza virus type A using reverse transcription loop-mediated isothermal amplification assisted with rod-shaped gold nanoparticles

Abstract

Reverse transcription loop-mediated isothermal amplification (RT-LAMP) and rod-shaped gold nanoparticles (gold nanorods; GNRs) were employed for nanomolecular detection of human influenza virus type A RNA. After cDNA synthesis from the RNA, the primers targeting the M protein gene were used for LAMP amplification. A blue shift from red to purple from the GNR inserting into the LAMP–DNAs can be seen by the naked eye. Transmission electron microscopy revealed the formation GNR aggregates due to their interactions with LAMP DNA. One pg RNA (10⁻³ dilution of the viral cDNA) was detected using this colorimetric test. The nanomolecular test showed 100% sensitivity and 95.8% specificity in comparison to results by RT-PCR. Also, the test indicated 100% sensitivity and 100% specificity in comparison to results by RT-LAMP. The described nanomolecular test could detect human influenza virus type A RNA in nearly 1 hour.