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.
Original Research
6.
Real time RT-PCR assay for detection of different
serotypes of FMDV in Egypt - Laila El-Shehawy, A. M. H. Azab, W. Mossad, Ehab El-Sayed,
A. Ismail, Wafaa Deghady
Vet World. 2012; 5(12): 732-737
doi:
10.5455/vetworld.2012.7
32-737
Abstract
Aim:
The present study indicated that rRT-PCR could be
provided for the detection of FMDV in infected,
contact and carrier cattle and also provide a rapid
sensitive tool aiming to aid in rapid disease
detection and control. Foot and Mouth disease virus
serotypes O1 and A still existing in Egypt. In
January 2012, sever outbreaks struck the animal
population in most Egyptian governorates. The
causative virus was identified as FMDV SAT2.
Material and Methods: Five samples of tongue
epithelium (ET) and five oesophageal-pharyngeal (OP)
fluid samples were collected from FMD suspected
cattle in infected farm at El-Fayoum and 20 OP
samples from in-contact cattle at the same farm in
addition to 30 OP samples from apparently healthy
cattle at three different localities in El-Fayoum
governorate (12 from Fayoum; 9 from Sinoras and 9
from Edsa) in order to detect carrier cattle. All of
these samples were collected during November and
December 2011 and January 2012.
Results: All
the ET and OP samples were inoculated on BHK cell
culture and baby mice. The obtained results were
identified using complement fixation test in
addition to real-time reverse transcriptase
polymerase chain reaction (rRT-PCR). In the infected
farm at El-Fayoum FMDV type SAT2 was detected in
cattle which are considered as the first
introduction of this type while FMDV type O and SAT2
were detected in the in-contact cattle in the same
farm. The sensitivity of rRT-PCR was cleared in the
in-contact cattle as 13 out of 20 OP samples were
positive to FMDV by rRT-PCR while 11 out of 20 OP
samples were positive to FMDV by CFT. The OP samples
collected from apparently healthy cattle from Fayoum,
Sinoras and Edsa localities in Fayoum governorate
demonstrate the circulation of the FMDV type A, O
and the recent SAT2 in carrier cattle which threaten
cattle population in Fayoum governorate. Also the
sensitivity of real time RT-PCR over the CFT in
detection of FMDV carrier cattle was clearly noticed
in these localities as 19 out of 30 OP samples were
positive by rRT-PCR while in contrast there were
only 16 out of 30 samples positive by CFT.
Conclusion: In conclusion, this study
demonstrates that real-time RT-PCR currently used at
the WRL for FMD provides an extremely sensitive and
rapid additional procedure for improved laboratory
diagnosis of FMD especially in-contact and carrier
cases. The rRT-PCR generated results in less than
one day from test commencement, in contrast to up to
four days to define some positive and all negative
samples by combined use of CFT and virus isolation.
This is an important feature when definitive
diagnostic results are required in a short timescale
during emergencies. Also this study demonstrates the
current situation of FMDV circulating in EL-Fayoum
governorate and the introduction of new SAT2
serotype beside type A and O.
Keywords: FMDV,
Typing, Isolation, OP, ET, SAT-2, rRT-PCR, Carrier,
Cattle
