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. This journal is © the Partner Organisations 2014