Marquardt O, Ohlinger V F
Bundesforschungsanstalt für Viruskrankheiten der Tiere, Tubingen, Germany.
J Virol Methods. 1995 Jun;53(2-3):189-99. doi: 10.1016/0166-0934(95)00014-l.
Monoclonal antibodies directed against an isolate of swine vesicular disease virus (SVDV), characterized by virus neutralization tests and competition assays, were used to compare SVDV isolates and isolates of the antigenically related Coxsackie viruses by ELISA. SVDV-specific reaction patterns and one specific for Coxsackie viruses were observed. This provided a method for distinguishing between these enteroviruses. In addition, RT-PCRs were undertaken with Coxsackie virus and SVDV genomes. Different product patterns were obtained which correlated with the genetic differences revealed by nucleotide sequence determination. RT-PCR distinguished between SVDV and Coxsackie viruses by pattern differences. Further SVDV-specific PCRs were carried out with clinical samples. Viral genomes were detected with a sensitivity equivalent to that of virus isolation in cell culture. Sequencing of the Coxsackie virus-derived 2A-coding PCR products resulted in a not previously described sequence of a B5 isolate and in SVDV-specific sequence of two Coxsackie virus A16 isolates. The differences of the isolates by ELISA and PCR reactivity, as well as the nucleotide sequence differences are consistent with the quasispecies concept of RNA viruses.
通过病毒中和试验和竞争试验对针对猪水疱病病毒（SVDV）分离株的单克隆抗体进行了表征，并用酶联免疫吸附测定法（ELISA）比较了SVDV分离株与抗原相关的柯萨奇病毒分离株。观察到了SVDV特异性反应模式和一种柯萨奇病毒特异性反应模式。这提供了一种区分这些肠道病毒的方法。此外，对柯萨奇病毒和SVDV基因组进行了逆转录聚合酶链反应（RT-PCR）。获得了不同的产物模式，这些模式与通过核苷酸序列测定揭示的遗传差异相关。RT-PCR通过模式差异区分了SVDV和柯萨奇病毒。对临床样本进行了进一步的SVDV特异性PCR。检测到病毒基因组，其灵敏度与细胞培养中的病毒分离相当。对源自柯萨奇病毒的2A编码PCR产物进行测序，得到了一个先前未描述的B5分离株序列和两个柯萨奇病毒A16分离株的SVDV特异性序列。分离株在ELISA和PCR反应性方面的差异以及核苷酸序列差异与RNA病毒的准种概念一致。
