Figure 4: Four-plex pooling of saliva specimens —

Four-plex pools were generated with saliva samples previously determined to be positive or negative for the presence of SARS-CoV-2 RNA. 81 wells were generated to contain four negative samples and eight contained one positive sample mixed with 3 negative samples. a, All wells containing positive sample pools were called either positive (7/8) or inconclusive (1/8). b, Viral and MS2 Ct values for unpooled and pooled positive saliva samples. Note that unpooled saliva sample 2 was run in two separate positive pools while all other positive pools contained unique positive samples. Samples were sorted by the N gene Ct value in unpooled saliva samples. Ct values >37 are shaded in gray. Undetected Ct values are plotted as zero and designated by “ND”, not detected. c, Results of pooled samples are compared to the final results for unpooled saliva samples run on the finalized saliva extraction protocol. “Specimen Insufficient” pooled samples were subsequently re-run either in new pools or unpooled to obtain a final result.