Multiplex logic processing isothermal diagnostic assays for an evolving virus

Sanchita Bhadra; Miguel A. Saldaña; Hannah Grace Han; Grant L. Hughes; Andrew D. Ellington

Capsid and NS1 LAMP-OSD assays and 4-Plex-LAMP-2GO assays of the remaining half of the ZIKV-fed mosquitoes remained as dark as primer-less assays (Supplementary Figure 2) . Our LAMP-based assays were in 100% agreement with TaqMan qRT-PCR, which also failed to detect ZIKV nucleic acids in these ZIKV-fed mosquitoes (Figure 8) . These results suggest an absence of detectable infection in these mosquitoes, which is consistent with previously reported infection rates. 34, 35, 36, 37, 38 Specificity of LAMP-based assays was confirmed by analyzing 6 additional age-matched Ae. aegypti mosquitoes that had not received a ZIKVspiked blood meal and were negative for ZIKV when tested by qRT-PCR. None of the uninfected mosquitoes triggered a false positive signal; LAMP assays containing these uninfected mosquitoes remained indistinguishable from primer-less assays (Supplementary Figure 2) .