Rhesus macaques can be effectively infected with SARS-CoV-2 via ocular conjunctival route

Wei Deng; Linlin Bao; Hong Gao; Zhiguang Xiang; Yajin Qu; Zhiqi Song; Shunran Gong; Jiayi Liu; Jiangning Liu; Pin Yu; Feifei Qi; Yanfeng Xu; Fengli Li; Chong Xiao; Qi Lv; Jing Xue; Qiang Wei; Mingya Liu; Guanpeng Wang; Shunyi Wang; Haisheng Yu; Xing Liu; Wenjie Zhao; Yunlin Han; Chuan Qin

The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.13.990036 doi: bioRxiv preprint log10 RNA copies/mL) lymph node, stomach (4.61 log10 RNA copies/mL), duodenum 85 (4.66 log10 RNA copies/mL), ileum (1.08 log10 RNA copies/mL) and caecum (1.06 86 log10 RNA copies/mL) (Figure 1d ). By contrast, for IT-1, the distribution of virus 87 might be somewhat different owing to viral replication was highly in different lobes 88 of the lung (10 4.22 to 10 7.81 copies/mL), and viral load was also widely detected in the 89 nasal septum (4.69 log10 RNA copies/mL), tracheas (6.61 log10 RNA copies/mL), The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.13.990036 doi: bioRxiv preprint The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.13.990036 doi: bioRxiv preprint An electric homogenizer was applied to prepare tissues homogenates by 2 min 30s 221 incubated in 1ml of DMEM. The homogenates were centrifuged at 3,000 rpm at 4°C 222 for 10 min. The supernatant was harvested and reposited for viral titer at −80°C. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.13.990036 doi: bioRxiv preprint sections at 37°C for 1 min. Three percent H2O2 in methanoland were fulfilled to 246 quench for endogenous peroxidases for 10 min. The slices were incubated at 4℃ 247 overnight with a laboratory prepared-7D2 monoclonal antibody after blocking in one 248 percent normal goat serum. HRP-labeled goat anti-mouse IgG secondary antibody 249 from ZDR-5307, Beijing ZSGB Biotechnology were maintained at 37°C for 60 min. 250 The 3,30-diaminobenzidine tetrahydrochloride was treated to make the results visual. 251 The sections were counterstained with hematoxylin, dehydrated and mounted on a The darker the blue color, the higher the viral load. The specific IgG antibody against 305 SARS-CoV-2 in the sera of the inoculated-macaques were tested by ELISA on 0, 7, 306 14, and 21 dpi (E). According to unpaired Welch's t-test, the specific IgG antibody in 307 the sera of conjunctival inoculated macaque exhibited a significant increase compared 308 with prior to inoculation (**p=0.0027) and 21 dpi (**p=0.0039). CJ-1 and CJ-2 were 309 the two macaques that inoculated via conjunctival route, IT-1 was the macaque that 310 inoculated via intratracheal route. IG-1 and IG-2 were the two macaques that 311 author/funder. All rights reserved. No reuse allowed without permission.