Epizootic hemorrhagic disease: Analysis of tissues by amplification and in situ hybridization reveals widespread orbivirus infection at low copy numbers
Epizootic hemorrhagic disease: Analysis of tissues by amplification and in situ hybridization reveals widespread orbivirus infection at low copy numbers
复制标题
DOI：
10.1128/jvi.72.5.3863-3871.1998
复制
发表时间：
1998-05-01
影响因子：
5.4
通讯作者：
Wilson, WC
中科院分区：
文献类型：
--
作者：
Brodie, SJ;Bardsley, KD;Wilson, WC
最近在美国西北部的野生反刍动物中爆发的出血热的特征是快速发热，随后不久出血和死亡。结果，在3个月的时间里，确认有1,000只白尾鹿和叉角羚羊死亡。病变是多系统的，包括重度水肿、充血、急性血管坏死和出血，具有与出血热一致的临床体征和/或病变的死亡动物通过放射免疫沉淀法具有动物流行性出血性疾病病毒血清型2（EHDV-2）抗体，但抗体仅限于免疫球蛋白M类。这些发现表明存在急性感染，并通过观察到大量鹿死亡得到证实;然而，在疫情期间很少看到临床受影响的鹿。此外，仅在出血性病变的动物中分离出EHDV-2和/或通过病毒型特异性逆转录（RT）-PCR检测到红细胞相关的EHDV-2 RNA。通过使用新的RT原位PCR检测，病毒核酸定位于出血组织中大量组织白细胞和血管内皮细胞的细胞质和血管，表明急性内膜和中膜坏死。由于原位杂交前的PCR扩增对于检测EHDV是必不可少的，单个细胞内的病毒拷贝数很低，
A recent outbreak of hemorrhagic fever in wild ruminants in the northwest United States was characterized by rapid onset of fever, followed shortly thereafter by hemorrhage and death. As a result, a confirmed 1,000 white-tailed deer and pronghorn antelope died over the course of 3 months. Lesions were multisystemic and included severe edema, congestion, acute vascular necrosis, and hemorrhage, Animals that died with clinical signs and/or lesions consistent with hemorrhagic fever had antibody to epizootic hemorrhagic disease virus serotype 2 (EHDV-2) by radioimmune precipitation but the antibody was limited exclusively to class immunoglobulin M. These findings, indicative of acute infection, were corroborated by the observation that numerous deer were found dead; however, clinically affected deer were rarely seen during the outbreak. Furthermore, only in animals with hemorrhagic lesions was EHDV-2 isolated and/or erythrocyte-associated EHDV-2 RNA detected by serotype-specific reverse transcription (RT)-PCR, By using a novel RT in situ PCR assay, viral nucleic acid was localized to the cytoplasm of large numbers of tissue leukocytes and vascular endothelium in tissues with hemorrhage and to vessels, demonstrating acute intimal and medial necrosis. Because PCR amplification prior to in situ hybridization was essential for detecting EHDV, the virus copy number within individual cells was low,