Respiratory picornavirus genotyping conventional nested RT-PCR ("Wisdom VP42 assay") v2

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This is my preferred, previously published [Ref 1], rhinovirus (RV) and enterovirus (EV) genotypingassay when seeking to identify the genotype of a respiratory picornavirus detected in a clinical sample extract. It is employed after use of a screeningreal-timeRT-PCR has identified a respiratory picornavirus. I have not confirmedthat it can detectevery single RV genotype but I do know that it detectsmanyfrom each of the threeRV species (Human rhinovirus A,Human rhinovirus BandHuman rhinovirus C) as well as at least someHuman enterovirus(EV) genotypes. The assay picks up EVs due to the shared genetic similarities inthe5'UTR target region. EVs can be discriminatedusing subgenomic sequencing (see VP42 typing assay protocol), or simply described as 'respiratory EVs' since there is no specific-specific vaccine ortreatment available anyway. This is arobust primary subgenomic sequencing assay. It is more sensitive than any VP1 protocols because it targets more conserved primer target sites. It produces a more reliabletyping result than does the 5'UTR region alone.

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