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Semi-quantitative digital analysis of polymerase chain reaction-electrophoresis gel: Potential applications in
John F Antiabong1, Mafora G Ngoepe2, Adakole S Abechi3
1Department of Molecular Microbiology, School of Biological Sciences, Flinders University SA, Bedford Park, 5042, Australia; Department of Applied Biotechnology, National Veterinary Research Institute, Nigeria.
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.
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