Kinetics raw dataMolecular and functional disparities of bovine viral diarrhoea virus 1b, 1d, and 1f subgenotype field isolates

Non-cytopathogenic (ncp) and cytopathogenic (cp) pairs of subgenotype 1b and 1f, and a single ncp isolate of the 1d subgenotype isolate of bovine viral diarrhoea virus (BVDV) strains currently circulating in Hungary were submitted to a comparative analysis regarding their <i>in vitro</i> replication kinetics and molecular composition. The kinetic analysis included digital PCR (dPCR) and virus titration. While the dPCR indicated a higher replication ratio for the cp viruses over the course of the 120 hrs observation period, the opposite was seen for all but the 1f ncp isolate by virus titration.The 1d/1b, 1d/1f, and 1b/1f subgenotypes differed in 431-510 amino acids on the polyprotein level. The 1b and 1f ncp/cp counterparts differed only in 9 and 29 amino acids, respectively. Subgenotype specific differences were also revealed in both untranslated regions.A „hallmark” amino acid deletion in the nucleocapsid coding region of the 1d subgenotype was present in the analysed isolate.None of the reported indels responsible for the ncp to cp transversion were identified in the respective pairs, however, amino acid substitutions were found in the Npro, capsid, and NS4B coding regions.Our results demonstrated a subtle but remarkable difference between the detection methods used for kinetic analysis, which can even be more pronounced <i>in vivo</i>, such as evaluating virus shedding results.Further, our findings support the previous observations regarding the complexity of the molecular differences between the biotypes of BVDV.针对目前在匈牙利流行的牛病毒性腹泻病毒（bovine viral diarrhoea virus，BVDV）毒株，本研究选取了1b和1f亚型的非致细胞病变型（non-cytopathogenic，ncp）与致细胞病变型（cytopathogenic，cp）配对株，以及1d亚型的1株单独非致细胞病变型分离株，对其体外（in vitro）复制动力学与分子组成开展比较分析。本次动力学分析包含数字PCR（digital PCR，dPCR）与病毒滴定实验。在120小时的观察周期内，数字PCR结果显示致细胞病变型毒株的复制水平更高；但通过病毒滴定实验检测，除1f亚型的非致细胞病变型分离株外，其余毒株的检测结果均与之相反。1d/1b、1d/1f及1b/1f亚型组合在多聚蛋白水平上存在431~510个氨基酸差异。1b与1f亚型的非致/致细胞病变型对应毒株之间，仅分别存在9个和29个氨基酸差异。在两个非翻译区（untranslated region，UTR）中也发现了亚型特异性差异。在所分析的1d亚型分离株的核衣壳编码区中，存在一处标志性氨基酸缺失。在对应的毒株配对中，未发现任何已报道的、可介导非致细胞病变型向致细胞病变型转变的插入缺失突变；但在Npro、衣壳蛋白及NS4B编码区中检测到了氨基酸替换。本研究结果表明，用于动力学分析的两种检测方法之间存在细微但显著的差异，这种差异在体内（in vivo）环境中甚至会更为突出，例如在评估病毒排毒结果时。此外，本研究结果佐证了此前关于牛病毒性腹泻病毒不同生物型间分子差异复杂性的相关研究结论。