Optimization of the Amplicons Detection System of Loop-Mediated Isothermal Amplification on Microfluidic Compact Disk
Salmonellosis caused by the bacteria in the genus Salmonella remains the most important foodborne disease in both developing and developed countries. Since several decades various microbial detection and quantification methods have been developed. One of recently developed gene amplification method is loop-mediated isothermal amplification (LAMP) which has been developed on tube-based platform. To facilitate the detection of pathogenic diseases in remote areas, a microfluidic platform has been developed called lab-on-a-CD. This paper presents an improved endpoint detection system for LAMP on Lab-on-a-CD platform. A set of ultraviolet (UV) emitter and color sensor have been used in this detection system to detect the emission level of LAMP amplicons. A LAMP assay has been performed with Salmonella bacteria DNA and calibrated the duty cycle of UV emitter to optimize the amplicons detection system.
KeywordsPathogens Salmonella Food safety Diagnosis Loop-mediated isothermal amplification LAMP Microfluidics Compact disc Lab-on-a-CD
We would like to thank Koh Xiu Pei from Microbiology Unit, Institute of Biological Sciences, Faculty of Science, University of Malaya for the assistance to conduct LAMP assay in tube-based method.
This work is financially supported by University of Malaya (UM) research grant, UM Research Collaborative Grant Scheme (Project No: CG003-2013) and MOSTI Science Fund (Project No: GA013-2013).
- 4.A. K. Bhunia, “Biosensors and bio‐based methods for the separation and detection of foodborne pathogens,” Advances in food and nutrition research, vol. 54, pp. 1–44, 2008.Google Scholar
- 6.G. Jeníková, J. Pazlarová, and K. Demnerová, “Detection of Salmonella in food samples by the combination of immunomagnetic separation and PCR assay,” International Microbiology, vol. 3, pp. 225–229, 2010.Google Scholar
- 15.K. T. Lim, C. S. J. Teh, and K. L. Thong, “Loop-mediated isothermal amplification assay for the rapid detection of Staphylococcus aureus,” BioMed research international, vol. 2013, 2013.Google Scholar
- 16.C. S. J. Teh, K. H. Chua, Y. A. L. Lim, S. C. Lee, and K. L. Thong, “Loop-Mediated Isothermal Amplification Assay for Detection of Generic and Verocytotoxin-Producing Escherichia coli among Indigenous Individuals in Malaysia,” The Scientific World Journal, vol. 2014, 2014.Google Scholar
- 17.(13 Nov 2014). PrimerExplore V4. Available: http://primerexplorer.jp/elamp4.0.0/index.html.