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Research
(Published
online: 27-10-2014)
23. Diagnosis of bovine foot and mouth
disease virus by real-time polymerase chain reaction and
nucleotide sequencing from outbreak herd samples in Ilesha Baruba,
Kwara state, Nigeria - Olatunde Hamza Olabode, Haruna
Makajuola Kazeem and Mashood Abiola Raji
Veterinary World, 7(10): 868-875
doi:
10.14202/vetworld.2014.868-875
Olatunde Hamza Olabode:
Department of Veterinary Microbiology, Faculty of Veterinary
Medicine, University of Abuja, Abuja, Nigeria; Department of
Veterinary Microbiology, Faculty of Veterinary Medicine, Ahmadu
Bello University, Zaria, Nigeria;
olabodeok@yahoo.com
Haruna Makajuola Kazeem:
Department of Veterinary Microbiology, Faculty of Veterinary
Medicine, Ahmadu Bello University, Zaria, Nigeria;
haruna_kazeem@yahoo.com
Mashood Abiola Raji: Department of Veterinary Microbiology,
Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria,
Nigeria;
rajmash2002@gmail.com
Received: 23-05-2014, Revised: 06-09-2014, Accepted: 10-09-2014,
Published online: 27-10-2014
Corresponding author:
Olatunde
Hamza Olabode, e-mail: olabodeok@yahoo.com
Abstract
Aim:
Molecular diagnosis of bovine foot and mouth disease virus (FMDV)
from outbreak herd in Bukaru-Rontuwa, Sinawu/Tumbunya ward of
Ilesha Baruba, in Kwara state-Nigeria was conducted to establish
the associated serotypes and disease control plan.
Materials and Methods: Purposive study was conducted in cattle
outbreak herds during the dry season of January-March, 2011.
Random sampling of blood and observed epithelial tissues was
collected, stored in accordance with standard methods and
subjected to RNA extraction and real-time reverse transcription
polymerase chain reaction (rRT-PCR). Positive samples for FMDV
were further subjected to reverse transcription polymerase chain
reaction (RT-PCR), nucleotide sequencing using sequence primers of
serotypes O, A, SAT 1-3 and gel electrophoresis. Obtained data
were interpreted based on NCBI BLASTN program.
Results: Foot and mouth disease (FMD)-RNA extract was not
found in all the blood tested with beta-actin range of Ct = 30-34.
rRT-PCR assay showed two positive samples with Ct values of 18.79
and 15.28. Gel electrophoresis identified sequenced PCR amplicons
as serotype A and SAT 2 respectively. Direct product sequencing
confirmed SAT 2 serotype was closely related to SAT 2 isolate
LIB/7/2003. Cloned RT-PCR product in pGEM-T easy vector confirmed
serotype A as closely related to sequence of A/NIG/21/2009, though
multiple NIG/2009 sequences were also identified as closely
related. Both isolates showed marked genetic homogeneity with >93%
genetic identity in the VP1 region which confirmed heterogeneity
and antigenic variation nature of FMDV.
Conclusion: Quasi species and subtypes of FMD serotypes A and
SAT 2 similar to A/NIG/21/2009 and SAT 2/LIB/7/2003 respectively
caused the reported FMD outbreaks in Fulani livestock herds
investigated. A combined real-time and optimized RT-PCR protocols
that would facilitate effective and timely FMD outbreak control
plan based on identified serotypes is thus suggested.
Keywords: foot and mouth disease virus, Ilesha Baruba, Kwara
State, molecular, outbreaks, phylogenetic.
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