Fig 1: Analytical performance comparison between Trioplex assay multiplex and the ZIKV singleplex format assay using small volume and large volume RNA extraction. Normal human serum or urine was contrived with ZIKV at a dilution of 1:10 before the limit of detection (1:10 BLoD), at the limit of detection (LoD), and at 1:10 after the limit of detection (1:10 ALoD). Twenty replicates of every dilution were extracted using the MagNA Pure 96 instrument (Roche) and tested by Trioplex assay multiplex or ZIKV singleplex format assay on the ABI 7500 Fast Dx or the QuantStudio Dx instruments. a Compares the mean genome copy equivalents per PCR reaction (GCE/rxn) of viral RNA extracted from serum at each dilution on the ABI 7500 Fast Dx instrument. A linear regression was plotted for multiplex with small volume protocol (Sv) (0.2 mL) (black straight line), singleplex assay with small volume protocol (gray straight line), multiplex with large volume protocol (Lv) (1 mL) (black dashed line). and singleplex assay with large volume protocol (gray dashed line). b Compares the mean genome copy equivalents per PCR reaction (GCE/rxn) of viral RNA extracted from serum or urine at each dilution on the QuantStudio Dx instrument. A linear regression was plotted for multiplex with small volume protocol serum (Sv) (black straight line), multiplex with large volume protocol serum (Lv) (gray straight line), multiplex with small volume protocol urine (Lv) (black dashed line), and multiplex with large volume protocol urine (gray dashed line). Error bars represent GCE/mL standard deviation. The CT values for every dilution replicate in serum tested was plotted for c small volume and d large volume extractions