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Open Access
Copyright: The authors. This article is an open access
article licensed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/2.0) which permits unrestricted use,
distribution and reproduction in any medium, provided the work is properly
cited.
Research
(Published
online: 13-02-2014)
5.
Molecular identification of local field
isolated fowl pox virus strain from Giza governorate of Egypt - Susan S. El-Mahdy, M. H.
H. Awaad and Y. A. Soliman
Veterinary World, 7(2): 66-71
doi:
10.14202/vetworld.2014.66-71
Abstract
Aim: Molecular identification
of field isolated pox virus from infected chickens in Egyptian
farms in 2012 by polymerase chain reaction (PCR).
Materials and Methods: Isolation and identification of fowl
pox virus (FPV isolate ch-08TK) from 30 day-old chickens
manifesting pox lesions. The isolate was propagated successfully
on chorioallantioc membrane of specific pathogen free (SPF)
embryonated chicken eggs and clear pock lesions were observed.
These lesions were homogenated and used to infected SPF chickens
and pigeons (via wing web route for chickens and via feather
follicle route into the thigh of pigeons) using 104.5 EID50/mL;
uninoculated birds of each species were used as negative controls
for determining the effect of isolated FPV strain.
Result: The isolated strain gave pathogenic takes 100% in
chickens and 75% in pigeons. Virus identification by PCR was done
using dream Tag master mix kit using the primers that targeted
thymidinekinase (TK) gene. These primers were designed using
Lasergene DNASTAR software Version 10. We used these primers to
amplify a 305 bp fragment of the TK gene of FPV. Phylogenetic
analysis of sequenced TK amplicone which reflects a new emerging
isolate of the field isolated FPV gave very limited similarity
(not exceeding 60%) with the published sequences. Thus FPV isolate
ch-08 TK gene (with Accession No. KF314718 in Gen Bank) is
different than canary, Egypt, 2012, P4b and Elsharqyia-FWPV2
FPV140 (FPV140); TKPV FPV140 (FPV140) and PGPV FPV (FPV140) which
have been isolated from cases of avipox virus in 2011 from skin
crust of different domesticated birds reared under the Egyptian
backyard management system. Our sequencing and phylogenetic
analysis of newly isolated virus using DNASTAR software Version 10
revealed that this virus differ from canary, Egypt, 2012, P4b and
Elsharqyia-(FWPV2 FPV140 (FPV140); TKPV FPV140 (FPV140) and PGPV
FPV (FPV140)) according to published data in Gen Bank.
Conclusion: FPV (isolated ch-08 TK gene with Accession
number KF314718 in Gen Bank) was isolated from 30 day old laying
chickens suffering from pox lesions. We believe that this study is
the first molecular identification of FPV strain from laying
chickens in Egypt.
Keywords: Egypt, fowl pox virus strain, local field
isolate, molecular identification.
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