Comparative growth kinetic of Egyptian LSDV isolate in Vero and MDBK cell line
Comparative growth kinetic of Egyptian LSDV isolate in Vero and MDBK cell line
Mohamed M. Mashaly1, Ayman H. El-Deeb2, Momtaz A. Shahein1, Hussein A. Hussein2
ABSTRACT
Background: Lumpy skin disease outbreaks were observed in different governorates of Egypt
affecting cattle throughout the year and are still present. The progressive shift of the outbreak in Egypt
during 2018 causing severe losses in cattle population despite vaccination, which may incur some
debates on the efficacy of current live attenuated vaccines against CaPVs. The virus isolation,
identification, and vaccine or other diagnostic preparations mainly depend on the proper culture used.
Objective: Our aim was to study the growth kinetic of recent LSDV isolate to determine the culture
properties of recent LSDV isolated from skin nodule that could be useful in the attempts to develop
more effective LSDV vaccine and diagnostic purposes.
Methods: In order to isolate the LSDV MDBK continuous cell line was used for six serial passages.
The obtained LSDV isolate was molecularly identified and deposited to the GenBank accession
number of MN271728. In addition, a comparative study of LSDV growth curves of two permissible
continuous cell lines (MDBK and Vero). The growth curve of LSDV was determined by inoculating
both cultures, harvesting at intervals up to 24 hours and titrating the virus culture. To confirm LSDV
accumulated in cell cultures, molecular identification is required; we carried Real-time PCR.
Results: This study has revealed that the virus culture period until 95% CPE was 48hr for MDBK and
72hr for Vero culture. The significant difference in growth behavior could be found between its growth
on MDBK and Vero cells that it reaches its highest peak at 48hr for MDBK and 72hr for Vero cells.
The maximum destruction in cell monolayers was observed within 72hr to 96hr post-infection. With an
optimal condition for both cultures, the LSDV titers were 5.2 in MDBK after 48hr and 5.67 in Vero
after 72hr. This rise in virus titer was followed by a sharp decrease after 48hr P.I. on MDBK unlike
Vero cells that continue its rise in virus titer until reaching its peak at 72hr P.I.
Conclusion: Comparison of growth kinetic of LSDV on MDBK and Vero cell line revealed that Vero
cell was considered the best susceptible cell for the propagation of LSDV with best harvesting time
72hr P.I to obtain a maximum titer for subsequent vaccine production.
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January 2020
Vol. 7, Pages 1-96
