OIE Reference Laboratory for VHS, National Fishery Products Quality Management Services, Busan 49111, Republic of Korea.
Department of Aquatic Life Medical Sciences, Sunmoon University, Asan-si, Chungnam 31460, Republic of Korea.
J Virol Methods. 2020 Feb;276:113733. doi: 10.1016/j.jviromet.2019.113733. Epub 2019 Sep 26.
Koi herpesvirus (KHV) disease is a serious disease in cultured carp (Cyprinus carpio). CCB and KF-1 cell lines are commonly used for virus isolation and observation of cytopathic effects (CPE) in carp and koi samples. The purpose of this study was to determine the optimal concentration of fetal bovine-serum (FBS) to use for supporting the replication of cyprinid herpesvirus 3 CyHV-3 in CCB and KF-1 cell lines. The following concentrations were tested: 0%, 2%, 5%, and 10% FBS. At 7 days post-viral inoculation (dpi), CPE with clear vacuolation was observed in both cell lines when supplemented with 0 and 2% FBS, but not in those supplemented with 5% or 10% FBS. At 14 dpi, CPE was observed in both cell lines supplemented with FBS at any of the tested concentrations when a high virus titer was inoculated. However, CPE was indistinguishable between cell lines supplemented with 10% FBS when a low virus titer was inoculated. Results of qPCR indicated that the number copies of the viral genome tended to be larger in both cell lines supplemented with 10% FBS than the corresponding number in cell lines supplemented with 0%, 2%, or 5% FBS, at 7 dpi. In conclusion, we recommend using 2% FBS as supplement for isolation and diagnosis of CyHV-3 viral infection in carp samples.
锦鲤疱疹病毒 (KHV) 病是养殖鲤鱼 (Cyprinus carpio) 的一种严重疾病。CCB 和 KF-1 细胞系常用于病毒分离和观察鲤鱼和锦鲤样本中的细胞病变效应 (CPE)。本研究旨在确定支持鲤鱼疱疹病毒 3 (CyHV-3) 在 CCB 和 KF-1 细胞系中复制的最佳胎牛血清 (FBS) 浓度。测试了以下浓度：0%、2%、5%和 10% FBS。在病毒接种后 7 天 (dpi)，当补充 0% 和 2% FBS 时，两种细胞系中均观察到具有清晰空泡的 CPE，但当补充 5%或 10% FBS 时则没有观察到。在接种高病毒滴度时，在任何测试浓度下补充 FBS 的两种细胞系中均观察到 CPE。然而，当接种低病毒滴度时，补充 10% FBS 的细胞系之间的 CPE 无法区分。qPCR 结果表明，在接种高病毒滴度时，与补充 0%、2%或 5% FBS 的细胞系相比，补充 10% FBS 的两种细胞系中的病毒基因组数量拷贝数在 7 dpi 时往往更大。总之，我们建议在鲤鱼样本中分离和诊断 CyHV-3 病毒感染时使用 2% FBS 作为补充。
