Aradaib I E, Mohammed M E, Mukhtar M M, Ghalib H W, Osburn B I
Department of Veterinary Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis 95616, U.S.A.
Comp Immunol Microbiol Infect Dis. 1997 Jun;20(3):211-8. doi: 10.1016/s0147-9571(97)00002-7.
The potential use of the recently reported polymerase chain reaction (PCR) protocol for detection of United States epizootic hemorrhagic disease virus (EHDV) serotype 1 (EHDV-1) and serotype 2 (EHDV-2) ribonucleic acid in cell culture and clinical specimens was evaluated for detection of Sudanese EHDV strains. EHDV serotype 5 (EHDV-5) and EHDV, isolate 318 (untyped) designated (EHDV-318), recovered from sentinel calves at the Khartoum University farm (Sudan) were studied. RNA from EHDV-5 and EHDV-318 and a number of EHDV field isolates, propagated in cell cultures, were detected by the described PCR-based assay. The specific 387 bp PCR products were visualized on ethidium-bromide stained agarose gel. Specificity of the PCR products was confirmed by chemiluminescent hybridization with non-radiolabeled internal probe. Amplification product was not detected when the PCR-based assay was applied to RNA from blutongue virus (BTV) prototypes serotypes 2, 10, 11, 13, 16 and 17; total nucleic acid extracts from uninfected BHK-21 cells. The results of this study indicated that the previously described EHDV-PCR assay could be applied for detection of Sudanese as well as United States strains of EHDV serogroup. In addition, the described EHDV-PCR assay could be used as a supportive diagnostic assay to the current conventional virus isolation procedures used for detection of EHDV infection in susceptible ruminants.
评估了最近报道的用于检测细胞培养物和临床标本中美国流行出血性疾病病毒1型（EHDV-1）和2型（EHDV-2）核糖核酸的聚合酶链反应（PCR）方案检测苏丹EHDV毒株的潜在用途。对从苏丹喀土穆大学农场的哨兵犊牛中分离出的EHDV 5型（EHDV-5）和EHDV分离株318（未分型，命名为EHDV-318）进行了研究。通过所述基于PCR的检测方法检测了在细胞培养物中繁殖的EHDV-5和EHDV-318以及一些EHDV野外分离株的RNA。在溴化乙锭染色的琼脂糖凝胶上观察到特异性的387 bp PCR产物。通过与非放射性标记的内部探针进行化学发光杂交，确认了PCR产物的特异性。当将基于PCR的检测方法应用于蓝舌病毒（BTV）2型、10型、11型、13型、16型和17型原型的RNA以及未感染的BHK-21细胞的总核酸提取物时，未检测到扩增产物。本研究结果表明，先前描述的EHDV-PCR检测方法可用于检测苏丹以及美国EHDV血清群毒株。此外，所述EHDV-PCR检测方法可作为目前用于检测易感反刍动物EHDV感染的传统病毒分离程序的辅助诊断方法。
