Haas L, Barrett T, Harder T, Bostock C J
Institute of Virology, Veterinary School Hannover, FRG.
Dtsch Tierarztl Wochenschr. 1990 Feb;97(2):93-5.
During the fatal seal epizootics in the North and Baltic Seas in summer 1988 a virus was isolated which was shown to be the causal agent. It was subsequently classified as morbillivirus by neutralization assays, reaction with monoclonal antibodies and nucleic acid hybridization studies. The virus (tentatively called Phocine Distemper Virus, PDV) is difficult to grow in culture making rapid diagnosis difficult. We have used the Polymerase Chain Reaction (PCR) as an alternative and fast method to detect the presence of virus-specific nucleic acid and we describe here the amplification of cell culture derived PDV RNA in a "one-tube" reaction using heterologous (Rinderpest Virus cDNA derived) F gene primers. The resulting 370 bp DNA fragment was shown to be morbillivirus derived by Southern blot hybridization using cloned RPV F gene as probe.
1988年夏季,北海和波罗的海发生了海豹致命性 epizootics,期间分离出一种病毒,该病毒被证明是病原体。随后通过中和试验、与单克隆抗体反应及核酸杂交研究,将其归类为麻疹病毒属病毒。该病毒(暂称为海豹瘟病毒,PDV)在培养中难以生长,使得快速诊断变得困难。我们使用聚合酶链反应(PCR)作为一种替代的快速方法来检测病毒特异性核酸的存在,并且在此描述了使用异源(源自牛瘟病毒cDNA)F基因引物在“单管”反应中扩增细胞培养物衍生的PDV RNA。使用克隆的RPV F基因作为探针,通过Southern印迹杂交表明所得到的370bp DNA片段源自麻疹病毒属病毒。