Formalin-fixed, paraffin-embedded tissues from European rabbits (Oryctolagus cuniculus) that succumbed to rabbit hemorrhagic disease virus 2 (RHDV2; Lagovirus GI.2) during the 2019 outbreak in Washington, USA, were utilized for in situ hybridization via RNAscope (ACDBio). This detection method was both sensitive and specific, with no staining in tissues from RHDV- (Lagovirus GI.1) and RHDV2-negative rabbits, and only slight background staining of RHDV-positive rabbits; RHDV2-positive tissues had bright-red cytoplasmic staining. Although much of the viral mRNA detection was consistent with previously described antigen detection via immunohistochemistry of the liver, lungs, and spleen, there was also significant glomerular staining in the kidneys, and endothelial staining within blood vessels of almost all organs. We validated the RNAscope technique for detection of RHDV2 mRNA in formalin-fixed, paraffin-embedded tissues, with increased sensitivity from previous techniques, and identified additional affected cell types that may contribute to the understanding of pathogenesis.

在 2019 年美国华盛顿爆发期间死于兔出血性疾病病毒 2 (RHDV2; Lagovirus GI.2) 的欧洲兔 ( Oryctolagus cuniculus )的福尔马林固定石蜡包埋组织被用于通过 RNAscope (ACDBio) 进行原位杂交）。这种检测方法既灵敏又特异，对 RHDV-（拉戈病毒）的组织没有染色。GI.1) 和 RHDV2 阴性兔，RHDV 阳性兔只有轻微的背景染色；RHDV2阳性组织细胞质呈鲜红色染色。虽然大部分病毒 mRNA 检测与之前描述的通过肝、肺和脾的免疫组织化学检测抗原一致，但在肾脏中也有显着的肾小球染色，并且在几乎所有器官的血管内都有内皮染色。我们验证了用于检测福尔马林固定、石蜡包埋组织中 RHDV2 mRNA 的 RNAscope 技术，与以前的技术相比具有更高的灵敏度，并确定了可能有助于了解发病机制的其他受影响的细胞类型。