Semi-quantitative digital analysis of polymerase chain reactionelectrophoresis gel: Potential applications in low-income veterinary laboratories

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

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 theFusobacterium 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.