Viruses, Vol. 13, Pages 1223: TASPERT: Target-Specific Reverse Transcript Pools to Improve HTS Plant Virus Diagnostics

Viruses, Vol. 13, Pages 1223: TASPERT: Target-Specific Reverse Transcript Pools to Improve HTS Plant Virus Diagnostics Viruses doi: 10.3390/v13071223 Authors: Andres S Espindola Daniela Sempertegui-Bayas Danny F Bravo-Padilla Viviana Freire-Zapata Francisco Ochoa-Corona Kitty F Cardwell High-throughput sequencing (HTS) is becoming the new norm of diagnostics in plant quarantine settings. HTS can be used to detect, in theory, all pathogens present in any given sample. The technique’s success depends on various factors, including methods for sample management/preparation and suitable bioinformatic analysis. The Limit of Detection (LoD) of HTS for plant diagnostic tests can be higher than that of PCR, increasing the risk of false negatives in the case of low titer of the target pathogen. Several solutions have been suggested, particularly for RNA viruses, including rRNA depletion of the host, dsRNA, and siRNA extractions, which increase the relative pathogen titer in a metagenomic sample. However, these solutions are costly and time-consuming. Here we present a faster and cost-effective alternative method with lower HTS-LoD similar to or lower than PCR. The technique is called TArget-SPecific Reverse Transcript (TASPERT) pool. It relies on pathogen-specific reverse primers, targeting all RNA viruses of interest, pooled and used in double-stranded cDNA synthesis. These reverse primers enrich the sample for only pathogens of interest. Evidence on how TASPERT is...
Source: Viruses - Category: Virology Authors: Tags: Article Source Type: research