EU and OIE Reference Laboratory for Classical Swine Fever, Institute of Virology, Department of Infectious Diseases, University of Veterinary Medicine, Hannover, Germany.
Vet Res. 2012 Jun 7;43(1):50. doi: 10.1186/1297-9716-43-50.
Molecular epidemiology has proven to be an essential tool in the control of classical swine fever (CSF) and its use has significantly increased during the past two decades. Phylogenetic analysis is a prerequisite for virus tracing and thus allows implementing more effective control measures. So far, fragments of the 5´NTR (150 nucleotides, nt) and the E2 gene (190 nt) have frequently been used for phylogenetic analyses. The short sequence lengths represent a limiting factor for differentiation of closely related isolates and also for confidence levels of proposed CSFV groups and subgroups. In this study, we used a set of 33 CSFV isolates in order to determine the nucleotide sequences of a 3508-3510 nt region within the 5´ terminal third of the viral genome. Including 22 additional sequences from GenBank database different regions of the genome, comprising the formerly used short 5´NTR and E2 fragments as well as the genomic regions encoding the individual viral proteins Npro, C, Erns, E1, and E2, were compared with respect to variability and suitability for phylogenetic analysis. Full-length E2 encoding sequences (1119 nt) proved to be most suitable for reliable and statistically significant phylogeny and analyses revealed results as good as obtained with the much longer entire 5´NTR-E2 sequences. This strategy is therefore recommended by the EU and OIE Reference Laboratory for CSF as it provides a solid and improved basis for CSFV molecular epidemiology. Finally, the power of this method is illustrated by the phylogenetic analysis of closely related CSFV isolates from a recent outbreak in Lithuania.
分子流行病学已被证明是控制经典猪瘟(CSF)的重要工具,在过去二十年中,其使用显著增加。系统进化分析是病毒溯源的前提,因此可以实施更有效的控制措施。到目前为止,5'NTR(150 个核苷酸,nt)和 E2 基因(190 nt)的片段经常用于系统进化分析。短序列长度是区分密切相关的分离株的一个限制因素,也是提出的 CSFv 组和亚组置信水平的限制因素。在本研究中,我们使用了一组 33 个 CSFv 分离株,以确定病毒基因组 5'端三分之一内的 3508-3510 nt 区域的核苷酸序列。包括来自 GenBank 数据库的 22 个额外序列,基因组的不同区域包括以前使用的短 5'NTR 和 E2 片段以及编码单个病毒蛋白 Npro、C、Erns、E1 和 E2 的基因组区域,这些序列在变异性和用于系统进化分析的适用性方面进行了比较。全长 E2 编码序列(1119 nt)被证明是最适合可靠和统计学上显著的系统发育和分析,结果与使用更长的整个 5'NTR-E2 序列获得的结果一样好。因此,该策略被欧盟和 OIE 参考实验室推荐用于 CSF,因为它为 CSFv 分子流行病学提供了一个坚实和改进的基础。最后,通过对立陶宛最近一次暴发中密切相关的 CSFv 分离株的系统进化分析,说明了该方法的有效性。