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Direct RT-PCR assay for virus detection and eriophyoid species identification

Tobiasz Druciarek: University of Arkansas

<div>Eriophyoid mites (Acari: Eriophyoidea) are important plant pests, causing direct damage and, most importantly vectoring plant viruses. Because of their size (averaging 200 µm in length) and concealed lifestyle (living in secreted or confined microhabitats) about 90% of eriophyoid mite fauna is speculated to be undiscovered. We have developed a novel direct Reverse Transcriptase - Polymerase Chain Reaction (direct RT-PCR) assays for species barcoding and virus detection in individual mites. Bypassed extraction steps prevent loss of nucleic acids during the extraction and lower the risk of cross-contamination. RT step not only allows for the detection of RNA viruses but also provides additional template of the fragments used for species barcoding (Cox1, 28S rRNA), enhancing the sensitivity and PCR yields when compared to other methods. Results indicate that direct RT-PCR serve as fast and cost effective method for virus detection and species identification using individual mites.</div>