Laboratory of functional and environmental ecology, Faculty of Sciences and Technology Sidi Mohammed Ben Abdellah University, Imouzzer Road, BP 2202 Fez, Morocco; Laboratory of Research and Development virology, MCI Animal Health, Lot. 157, Zone Industrielle Sud-Ouest (ERAC) B.P: 278, 28810 Mohammedia, Morocco.
Laboratory of Research and Development virology, MCI Animal Health, Lot. 157, Zone Industrielle Sud-Ouest (ERAC) B.P: 278, 28810 Mohammedia, Morocco.
J Virol Methods. 2021 Jul;293:114164. doi: 10.1016/j.jviromet.2021.114164. Epub 2021 Apr 14.
Lumpy skin disease virus (LSDV), sheeppox virus (SPPV) and goatpox (GTPV) virus have been usually grown on primary cells for diagnosis, production and titration purposes. The use of primary cells present several inconvenient, heavy preparation, heterogeneous cell population, non-reproducible viral titration and presence of potential endogenous contaminants. Therefore investigating sensitivity of candidate continuous cell lines is needed. In this study, we compared the above Capripox viruses (CaPVs) sensitivity of primary cells of four origin (heart, skin, testis and kidney), with three cell lines (Vero, OA3.Ts and ESH-L). We tested sensitivity for virus isolation, replication cycle and titration, revealed by cytopathic effect (CPE), immunoenzymatic staining and immunofluorescence. Our results show that ESH-L cells and primary fetal heart cells present the highest sensitivity for CaPVs growth and detection. Vero cells can replicate those viruses but without showing any CPE while the titer obtained on OA3.Ts is lower than primary and ESH-L cells. ESH-L cells are an effective alternative to primary cells use for growing Capripoxviruses and their diagnosis.
块状皮肤疾病病毒(LSDV)、绵羊痘病毒(SPPV)和山羊痘病毒(GTPV)通常在原代细胞上培养用于诊断、生产和滴定目的。使用原代细胞存在几个不便之处,如细胞准备繁琐、细胞群体异质性、不可复制的病毒滴定和潜在的内源性污染物。因此,需要研究候选连续细胞系的敏感性。在这项研究中,我们比较了来自四个来源(心脏、皮肤、睾丸和肾脏)的原代细胞和三种细胞系(Vero、OA3.Ts 和 ESH-L)对上述羊痘病毒(CaPVs)的敏感性。我们通过细胞病变效应(CPE)、免疫酶染色和免疫荧光检测,测试了病毒分离、复制周期和滴定的敏感性。我们的结果表明,ESH-L 细胞和原代胎牛心脏细胞对 CaPVs 的生长和检测具有最高的敏感性。Vero 细胞可以复制这些病毒,但没有显示任何 CPE,而在 OA3.Ts 上获得的滴度低于原代和 ESH-L 细胞。ESH-L 细胞是替代原代细胞用于生长羊痘病毒及其诊断的有效方法。