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Seminar: Application of a Carbon Nanotube Array for enrichment and increased detection by qPCR and NGS of TSWV in Nicotiana benthamiana and in situ detection from crude plant extracts using RT-RPA

Juan Francisco Iturralde Martinez, Graduate Student, Penn State
Juan Francisco Iturralde Martinez

Juan Francisco Iturralde Martinez

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September 16, 2019, 3:35 PM - 4:30 PM

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Carbon nanotubes have proven useful for a plethora of applications, among which, the entrapment and concentration of virus particles is one of the most remarkable from a plant pathology perspective. CNT-STEP is a carbon nanotube-based microfluidics device developed by researchers at PSU that we used to increase the limits of detection of Tomato spotted wilt orthotospovirus, one of the most damaging viruses in tomato production in Florida, Georgia and California. Our results show that carbon nanotubes can be used to directly visualize, by electron microscopy, virus particles trapped in the devices, without any extra preparation. Furthermore, we confirmed that CNT-STEP can be customized to deplete the plant host genome while increasing virus-derived reads, when CNT-STEPs are used in conjunction with high throughput sequencing. Additionally, a recombinase polymerase amplification was developed for detection of TSWV before the onset of infection. This RPA can be visualized in 3 different ways and attains qRT-PCR levels of sensitivity and specificity.  Overall, my work provides novel tools that will improve the likelihood to manage viral diseases in a timely and efficient manner, while decreasing the amount of pesticides traditionally used to control insect vectors of plant pathogens.