Comparison of Eleven RNA Extraction Methods for Poliovirus Direct Molecular Detection in Stool Samples

Figures

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  FIG 1. Evaluation of viral RNA recovery for PV positive stool samples by 11 extraction methods. (A and B) Box plots of cycle threshold (CT) value (A) and viral genome copies/μL (B) of the PV stool samples according to each extraction method. Extracted RNA was tested by RT-qPCR assay using PV specific primers and probe. Each gray circle represents a sample. Each stool sample was tested in the RT-qPCR in duplicates run in parallel with Sabin 1 standards. The box plots show the mean (represented by red dot), median, lower quartile, and upper quartile of the data points for samples extracted by each method, and the whiskers show the range. P values are based on a linear mixed effects regression analysis.

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  FIG 2. The proportion of PV positive samples recovered by the 11 extraction methods. The black bars indicate the 95% confidence intervals for the true proportion of samples extracted by each extraction method that were PV positive in the different PCR assays (nested VP1, PanEV RT-PCR, PanPV RT-PCR, and RT-qPCR).

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  FIG 3. The overall number of samples detected as PV positive and negative for the 11 extraction methods following combined detection with the different PCR assays. The percentage in the bars indicates the sensitivity result for each extraction method after combined PV detection using PV specific RT-qPCR, nested VP1 PCR, and PanEV and PanPV RT-PCR.

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  FIG 4. Plot of cost against time for the 11 extraction methods.