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R esearch
(Published online:
21-05-2015)
14.
Co-culture: A quick approach for isolation of street rabies virus
in murine neuroblastoma cells
-
A.
Sasikalaveni, K. G. Tirumurugaan, S. Manoharan, G. Dhinakar Raj
and K. Kumanan
Veterinary World, 8(5): 636-639
doi:
10.14202/vetworld.2015.636-639
A. Sasikalaveni:
Department of Animal Biotechnology, Madras Veterinary College,
Tamil Nadu Veterinary and Animal Sciences University, Chennai -
600 007, Tamil Nadu, India;
venisasikala@gmail.com
K. G. Tirumurugaan:
Department of Animal Biotechnology, Madras Veterinary College,
Tamil Nadu Veterinary and Animal Sciences University, Chennai -
600 007, Tamil Nadu, India;
tirumurugaankg@tanuvas.org.in
S. Manoharan:
Department of Animal Biotechnology, Madras Veterinary College,
Tamil Nadu Veterinary and Animal Sciences University, Chennai -
600 007, Tamil Nadu, India;
ulagaimano@yahoo.com
G. Dhinakar Raj:
Department of Animal Biotechnology, Madras Veterinary College,
Tamil Nadu Veterinary and Animal Sciences University, Chennai -
600 007, Tamil Nadu, India;
dirtrpvb@tanuvas.org.in
K. Kumanan: Dean,
Faculty of Basic Sciences, Tamil Nadu Veterinary and Animal
Sciences University, Madhavaram Milk Colony, Chennai - 600 051,
Tamil Nadu, India;
kumananrani@gmail.com
Received:
30-12-2014,
Revised:
14-04-2015, Acceped:
19-04-2015, Published
online:
21-05-2015
Corresponding author:
K. G. Tirumurugaan, e-mail: tirumurugaankg@tanuvas.org.in
Citation:
Sasikalaveni A, Tirumurugaan KG, Monoharan S, Dhinakar Raj G,
Kumanan K (2015) Co-culture: A quick approach for isolation of
street rabies virus in murine neuroblastoma cells, Veterinary
World 8(5):636-639.
Abstract
Background: Laboratory
detection of rabies in most cases is based on detection of the
antigen by fluorescent antibody test, however, in weak positive
cases confirmative laboratory diagnosis depends on widely accepted
mouse inoculation test. Cell lines like neuroblastoma have been
used to isolate the virus with greater success not only to target
for diagnosis, but also for molecular studies that determine the
epidemiology of the circulating street rabies strains and in
studies that look at the efficiency of the developed monoclonal
antibodies to neutralize the different rabies strains. Due to the
recent issues in obtaining ethical permission for mouse
experimentation, and also the passages required in the cell lines
to isolate the virus, we report herewith a co-culture protocol
using the murine neuroblastoma (MNA) cells, which enable quicker
isolation of street rabies virus with minimum passages.
Objective: This study is not to have an alternative diagnostic
assay, but an approach to produce sufficient amount of rabies
virus in minimum passages by a co-culture approach in MNA cells.
Materials and Methods: The MNA cells are co-cultured by
topping the normal cells with infected cells every 48 h and the
infectivity was followed up by performing direct
fluorescent-antibody test.
Results: The co-culture approach results in 100% infectivity
and hence the use of live mouse for experimentation could be
avoided.
Conclusion: Co-culture method provides an alternative for the
situations with limited sample volume and for the quicker
isolation of virus which warrants the wild type strains without
much modification.
Keywords: co-culture, isolation, fluorescent-antibody test,
murine neuroblastoma, rapid tissue culture infection test.
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