Veterinary World

     Open access and peer reviewed journal  

ISSN (Online): 2231-0916


Home l Editorial board l Instructions for authors l Reviewer guideline l Open access policy l Archives l FAQ

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;

Mafora G. Ngoepe: Onderstepoort Biological Products, Onderstepoort, Pretoria, 0110, South Africa;

Adakole S. Abechi: Department of Applied Biotechnology, National Veterinary Research Institute, Nigeria;


Received: 09-03-2016, Accepted: 28-07-2016, Published online: 03-09-2016


Corresponding author: John F. Antiabong, e-mail:

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.

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.

1. Nietfeld, J.C. (2010) Field necropsy techniques and proper specimen submission for investigation of emerging infectious diseases of food animals. Vet. Clin. North Am. Food Anim. Pract., 26(1): 1-13.
2. Van Guilder, H.D., Vrana, K.E. and Freeman, W.M. (2008) Twenty-five years of quantitative PCR for gene expression analysis. Biotechniques, 44: 619.
3. Abramoff, M.D., Magelhaes, P.J. and Ram, S.J. (2004) Image processing with Image. J. Biophotonics Int., 11: 36-42.
4. Berchtold, C.M., Wu, Z.H., Huang, T.T. and Miyamoto, S. (2007) Calcium-dependent regulation of NEMO nuclear export in response to genotoxic stimuli. Mol. Cell. Biol., 27: 497-509.
PMid:17074802 PMCid:PMC1800799
5. Rai, T., Mosoian, A. and Resh, M.D. (2010) Annexin 2 is not required for human immunodeficiency virus Type 1 particle production but plays a cell type-dependent role in regulating infectivity. J. Virol., 84: 9783-9792.
PMid:20631122 PMCid:PMC2937750
6. Antiabong, J.F., Boardman, W., Smith, I., Brown, M.H., Ball, A.S. and Goodman, A.E. (2013) A molecular survey of a captive wallaby population for periodontopathogens and the co-incidence of Fusobacterium necrophorum subspecies necrophorum with periodontal diseases. Vet. Microbiol., 63: 335-343.
7. Schmittgen, T.D., Zakrajsek, B.A., Mills, A.G., Gorn, V., Singer, M.J. and Reed, M.W. (2000) Quantitative reverse transcription–polymerase chain reaction to study mRNA decay: Comparison of endpoint and real-time methods. Anal. Biochem., 285: 194-204.
8. Cai, H.Y., Caswell, J.L. and Prescott, J. (2014) Nonculture molecular techniques for diagnosis of bacterial disease in animals a diagnostic laboratory perspective. Vet. Pathol. Online, 51(2): 341-350.
9. Sardelli, A.D. (1993) The plateau effect: Understanding PCR limitations. Amplif. Forum PCR Users, 9: 1-5.
10. McPherson, M.J., Hames, B.D. and Taylor, G.R. (1995) PCR: A Practical Approach. IRL Press, Oxford. p1-22.
11. Pfaffl, M.W. (2004) In: Bustin, S.A., editor. A-Z of Quantitative PCR. International University Line, La Jolla. p89-113.