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Complexities in Isolation and Purification of Multiple Viruses from Mixed Viral Infections: Viral Interference, Persistence and Exclusion

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Title Complexities in Isolation and Purification of Multiple Viruses from Mixed Viral Infections: Viral Interference, Persistence and Exclusion
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Creator Naveen Kumar, Sanjay Barua, Thachamvally Riyesh, Kundan K. Chaubey, Krishan Dutt Rawat, Nitin Khandelwal, Anil K. Mishra, Nitika Sharma, Surender S. Chandel, Shalini Sharma, Manoj K. Singh, Dinesh K. Sharma, Shoor V. Singh, Bhupendra N. Tripathi
 
Subject PPRV FMDV virus isolation complexities
 
Description Not Available
Successful purification of multiple viruses from mixed infections remains a challenge. In this
study, we investigated peste des petits ruminants virus (PPRV) and foot-and-mouth disease
virus (FMDV) mixed infection in goats. Rather than in a single cell type, cytopathic effect
(CPE) of the virus was observed in cocultured Vero/BHK-21 cells at 6th blind passage (BP).
PPRV, but not FMDV could be purified from the virus mixture by plaque assay. Viral RNA
(mixture) transfection in BHK-21 cells produced FMDV but not PPRV virions, a strategy which
we have successfully employed for the first time to eliminate the negative-stranded RNA virus
from the virus mixture. FMDV phenotypes, such as replication competent but noncytolytic,
cytolytic but defective in plaque formation and, cytolytic but defective in both plaque formation
and standard FMDV genome were observed respectively, at passage level BP8, BP15 and
BP19 and hence complicated virus isolation in the cell culture system. Mixed infection was
not found to induce any significant antigenic and genetic diversity in both PPRV and FMDV.
Further, we for the first time demonstrated the viral interference between PPRV and FMDV.
Prior transfection of PPRV RNA, but not Newcastle disease virus (NDV) and rotavirus RNA
resulted in reduced FMDV replication in BHK-21 cells suggesting that the PPRV RNAinduced
interference was specifically directed against FMDV. On long-termcoinfection of
some acute pathogenic viruses (all possible combinations of PPRV, FMDV, NDV and buffalopox
virus) in Vero cells, inmost cases, one of the coinfecting viruses was excluded at passage
level 5 suggesting that the long-term coinfection may modify viral persistence. To the
best of our knowledge, this is the first documented evidence describing a natural mixed infection
of FMDV and PPRV. The study not only provides simple and reliable methodologies for
isolation and purification of two epidemiologically and economically important groups of
viruses, but could also help in establishing better guidelines for trading animals that could
transmit further infections and epidemics in disease free nations.
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Date 2018-12-01T07:13:59Z
2018-12-01T07:13:59Z
2016-01-01
 
Type Article
 
Identifier doi:10.1371/journal.pone.0156110
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/14818
 
Language English
 
Relation Not Available;
 
Publisher PLOS