Forty-nine HCV E1E2 isolates show measurable infectivity in full-length chimeric HCV within 96 h of electroporation independent of function in a pseudoparticle assay. Focus forming units (FFU) were calculated from NS5a stained cells for each isolate at 24, 96 and 192 h time points. (a) Thirty-two genotype (Gt) 1 isolates, (b) 12 Gt2 isolates and (c) 5 Gt3 isolates are shown alongside the reference strains H77/JFH (Gt1, purple open circle), J6/JFH (Gt2, green open diamond) and JFH-1 (parental chimera; Gt2, green filled diamond). (d) FFU calculated for the 96 h time point for all 49 chimeras plotted against the isolate’s ability to produce infectious pseudoparticles above the limit of detection in a retroviral pseudotype assay (closed objects, termed pp +ve) or not (open objects, termed pp –ve). (e) Eight isolates with titres ranging from 1.2×103 to 1.1×106 FFU ml−1 were neutralized by an increasing concentration of anti-CD81 mAb. Mean±sem shown for each category.