Rapid Screening of Barley Stripe Mosaic Virus Via Microfluidic Chip Method

Abstract

<i>Barley stripe mosaic virus</i>, the type member of <i>Hordeivirus</i>, constitutes an important seed-transmitted pathogen distributed in almost every barley growing area worldwide. Researches on efficient detection of BSMV suitable in wild application are in urgent need. New platforms like microfluidic technology keep progressing rapidly in past decades and gains increasing influence on life sciences. Thus in this study, one rapid screening method for BSMV via microfluidic chip was established. Through method validity, we succeeded in demonstrating the amplification of BSMV within 30 min at one constant temperature using a microfluidic chip assay. Sensitivity analysis conformed that this method obtained a detection limit at 1.00×102 copies/μL, which was even more sensitive than real-time RT-PCR. <i>Maize chlorotic mottle virus</i>, <i>Maize dwarf mosaic virus</i>, <i>Oat mosaic virus</i>, and <i>Wheat streak mosaic virus</i> were chosen in specificity test along with BSMV, but only BSMV expressed typical amplification curves and Ct. Microfluidic analytic systems realize miniaturization by reducing the reaction volume to just 5 μL, which has several superiorities such as saving expensive reagents and reducing inspection costs. Moreover this microfluidic chip assay can simultaneously detect a variety of viruses, and the whole process takes just 1 hour, which greatly speeds up the detection and improves efficiency. All in all, this microfluidic chip method has the potential to be further implemented by phytosanitary services for routine diagnose as well as rapid screening in places like ports.