Madin-Darby bovine kidney (MDBK) cells are a suitable cell line for the propagation and study of the bovine poxvirus lumpy skin disease virus

Abstract

Highlights LSDV replicates to high titers (approximately 1 × 10 7 PFU/mL) in MDBK cells. LSDV forms foci-like poxviral plaques in MDBK cells. A plaque-reduction neutralisation test was developed for LSDV antibody quantitation. A method for purification of LSDV genomic DNA was optimized.

Isolation and characterization of lumpy skin disease virus from cattle in India

“…No CPE could be observed upon immediate infection of LSDV (isolated in PGK cells) to Vero cells. Later on, CPE characterized by cell clustering (foci), like those observed previously in MDBK cells [29] was observed at P4. However, virus did not form any plaques at this stage (P4).…”

“…However, primary cells are prone to contamination, time consuming and expensive to produce and are not considered appropriate with current efforts to reduce the use of animals in science. MDBK cells have been commonly used for the in vitro propagation of LSDV at high titres [ 28 , 29 ]. Some studies also suggest the use of MDBK cells for LSDV isolation [ 28 ] but it seems uncertain whether these are more sensitive than the primary cells for virus isolation.…”

“…A focus forming assay (in MDBK cells) has been described for LSDV [ 25 , 29 ] but precise counting of foci under microscope is a tedious task. Although MDBK cells are considered sensitive for in vitro propagation of LSDV [ 28 , 29 ], like in other cell types, virus does not produce plaques in MDBK cells. We for the first time adapted the LSDV in Vero cells by presuming that the LSDV may form plaques in an alternative cell line.…”

“…However, TCID 50 may not represent the accurate virus titre as manual observation of CPE under microscope could lead to deviations in the results partly due to the possible subjective (eye) effect. Emerging evidences also suggest use of MDBK cell line for propagation of LSDV [29][30][31][32]. A focus forming assay (in MDBK cells) has been described for LSDV [25,29] but precise counting of foci under microscope is a tedious task.…”

“…Emerging evidences also suggest use of MDBK cell line for propagation of LSDV [29][30][31][32]. A focus forming assay (in MDBK cells) has been described for LSDV [25,29] but precise counting of foci under microscope is a tedious task. Although MDBK cells are considered sensitive for in vitro propagation of LSDV [28,29], like in other cell types, virus does not produce plaques in MDBK cells.…”