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Research
(Published
online: 13-10-2014)
11.
Detection of African swine fever virus
from formalin fixed and non-fixed tissues by polymerase chain
reaction -
P. D. Luka, A. R.
Jambol and B. Yakubu
Veterinary World, 7(10): 811-815
doi:
10.14202/vetworld.2014.811-815
P. D.
Luka:
Applied Molecular Biology Department, National Veterinary Research
Institute, Vom, Plateau State, Nigeria; pamluka08@gmail.com
A. R.
Jambol:
Applied Molecular Biology Department, National Veterinary Research
Institute, Vom, Plateau State, Nigeria;
dtrufriend@yahoo.com
B.
Yakubu; Applied Molecular Biology Department, National
Veterinary Research Institute, Vom, Plateau State, Nigeria;
minitsayaks@yahoo.com
Received:
17-06-2014, Revised: 08-09-2014, Accepted: 11-09-2014, Published
online: 13-10-2014
Corresponding author:
P. D. Luka, e-mail: pamluka08@gmail.com
Abstract
Aim:
Formalin fixing and paraffin embedding of tissue samples is
one of the techniques for preserving the structural integrity of
cells for a very long time. However, extraction and analysis of
genomic material from formalin fixed tissue (FFT) remains a
challenge despite numerous attempts to develop a more effective
method. The success of polymerase chain reaction (PCR) depends on
the quality of DNA extract.
Materials and Methods: Here we assessed the conventional
method of DNA extraction from FFT for African swine fever virus
(ASFV) detection. The modified conventional method gave a
higher quality DNA when compared with commercially available DNA
extraction kits (QIAamp® DNA Mini Kit, DNeasy®
Blood and Tissue Kit, and ZR Genomic DNA™
Tissue MiniPrep).
Results: An average A260/A280 DNA purity of 0.86-1.68 and
3.22-5.32 μg DNA/mg for formalin fixed and non-fixed tissues,
respectively using a conventional method. In a reproducible and
three times repeat PCR, the ASFV DNA expected product size of 278
bp was obtained from the DNA extract of the conventional method
but not from the DNA extract of the commercial kits.
Conclusion: The present study has demonstrated that the
conventional method extracts ASFV genome better than commercial
kit. In summary, the commercial kit extraction appeared not
suitable to purify ASFV DNA from FFT. We, therefore, recommend
that the use of the conventional method be considered for African
swine fever DNA extraction from FFT.
Keyword: African swine fever, detection, formalin fixed,
non-formalin fixed, polymerase chain reaction.
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