Open Access
Research
(Published
online: 03-09-2016)
3.
Semi-quantitative digital analysis of
polymerase chain reactionelectrophoresis gel: Potential
applications in low-income veterinary laboratories -
John F. Antiabong, Mafora G. Ngoepe and Adakole S. Abechi
Veterinary World, 9(9): 935-939
doi:
10.14202/vetworld.2016.935-939
John F. Antiabong:
Department of Molecular Microbiology, School of Biological
Sciences, Flinders University SA, Bedford Park, 5042, Australia;
Department of Applied Biotechnology, National Veterinary
Research Institute, Nigeria; negribj@hotmail.com
Mafora G. Ngoepe:
Onderstepoort Biological Products, Onderstepoort, Pretoria,
0110, South Africa; ngoepeg@yahoo.com
Adakole S. Abechi:
Department of Applied Biotechnology, National Veterinary
Research Institute, Nigeria; jamush2002@yahoo.com
Received: 09-03-2016, Accepted: 28-07-2016, Published online:
03-09-2016
Corresponding author:
John F. Antiabong, e-mail: negribj@hotmail.com
Citation:
Antiabong JF, Ngoepe MG, Abechi AS (2016) Semiquantitative
digital analysis of polymerase chain reaction-electrophoresis
gel: Potential applications in low-income veterinary
laboratories,
Veterinary World, 9(9):
935-939.
Abstract
Aim:
The interpretation of conventional polymerase chain reaction (PCR)
assay results is often limited to either positive or negative
(non-detectable). The more robust quantitative PCR (qPCR) method
is mostly reserved for quantitation studies and not a readily
accessible technology in laboratories across developing nations.
The aim of this study was to evaluate a semi-quantitative method
for conventional PCR amplicons using digital image analysis of
electrophoretic gel. The potential applications are also
discussed.
Materials and Methods:
This study describes standard conditions for the digital image
analysis of PCR amplicons using the freely available ImageJ
software and confirmed using the qPCR assay.
Results and Conclusion:
Comparison of ImageJ analysis of PCR-electrophoresis gel and
qPCR methods showed similar trends in the
Fusobacterium necrophorum
DNA concentration associated with healthy and periodontal
disease infected wallabies (p≤0.03). Based on these empirical
data, this study adds descriptive attributes (“more” or “less”)
to the interpretation of conventional PCR results. The potential
applications in low-income veterinary laboratories are
suggested, and guidelines for the adoption of the method are
also highlighted.
Keywords:
applications, digital image analysis, ImageJ software,
polymerase chain reaction-electrophoresis, polymerase chain
reaction amplicon, quantitative polymerase chain reaction.
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