Analysis of the post-immunization serum neutralizing activity against ZIKV. The sera neutralizing activity was measured using a plaque reduction neutralization test (PRNT) against the following Zika virus strains: H/PAN/2016/BEI-259634 and MR766.

Analysis of the serum neutralizing activity against ZIKV. The sera neutralizing activity was measured using a plaque reduction neutralization test (PRNT) against the following Zika virus strains: H/PAN/2016/BEI-259634 and MR766.  Plaque reduction neutralization test (PRNT).To measure ZIKV neutralization by the obtained sera, PRNTs were performed. Briefly, 12-well cell culture plates were seeded with A549 cells to near confluence on the day of ZIKV infection. The sera were heat-inactivated at 56°C for 30 min. Sera were diluted 2-fold in fresh medium and mixed with equal volumes of ZIKV strain H/PAN/2016/BEI-259634 or MR766 containing approximately 50 PFU/well. The mixtures were incubated for 1 h at 37°C and used to infect cells. After 1 h, the inoculum was aspirated, and the cells were overlaid with 1% methylcellulose in MEM medium. The plates were incubated for 4 days at 37°C and stained with 0.5% crystal violet solution in 20% ethanol. Plaques were counted, and PRNT90 was estimated as the highest dilution that resulted in at least a 90% reduction in ZIKV plaques. The analysis was done twice. PRNT90 values estimated from each analysis are presented in the table.

抗寨卡病毒（Zika virus, ZIKV）血清中和活性分析。本研究采用空斑减少中和试验（plaque reduction neutralization test, PRNT），针对下述两株寨卡病毒毒株：H/PAN/2016/BEI-259634与MR766检测受试血清的中和活性。空斑减少中和试验（PRNT）：为测定所得血清对寨卡病毒的中和活性，本研究开展空斑减少中和试验。简言之，于寨卡病毒感染当日，将A549细胞接种于12孔细胞培养板，待细胞生长至接近汇合状态。将血清于56℃热灭活30分钟。将血清以新鲜培养基进行2倍梯度稀释，随后与等体积的寨卡病毒毒株H/PAN/2016/BEI-259634或MR766（每孔含约50个蚀斑形成单位（Plaque Forming Unit, PFU））混合。将混合液于37℃孵育1小时，随后用于感染细胞。孵育1小时后，吸弃接种培养液，向细胞层覆加含1%甲基纤维素的Minimum Essential Medium（MEM）培养基。将培养板置于37℃孵育4天，随后采用20%乙醇配制的0.5%结晶紫溶液进行染色。计数空斑数量，PRNT90值定义为可使寨卡病毒空斑数量减少至少90%的最高血清稀释度。本实验重复开展两次，两次分析所得的PRNT90值均列于表格中。