Paper-based visual detection of Salmonella bacteria using isothermal DNA amplification and magnetic bead aggregation
- Author:
Sharmili Roy, Sim Xiao Wei
;
Ibrahim Abd Rahman
;
Minhaz Uddin Ahmed
- Publication Type:Journal Article
- Keywords:
Food safety;
loop-mediated isothermal amplification;
magnetic beads;
Salmonella
- MeSH:
Salmonella
- From:Malaysian Journal of Microbiology
2016;12(5):332-338
- CountryMalaysia
- Language:English
-
Abstract:
Aims: The present study aimed to develop a new approach for detecting Salmonella species at picogram levels using
magnetic bead (MB) aggregation through loop-mediated isothermal amplification (LAMP).
Methodology and results: For the first time to our knowledge, Salmonella LAMP amplicons were analyzed using MB
aggregation. LAMPs were conducted with a simple heat block, and the results were compared with those obtained with
conventional LAMP-MB techniques. Furthermore, the volume and concentration of MB solutions were optimized. Our
method detected Salmonella genomic DNA at a low picogram level (1 pg/µL). The specificity of this method was also
examined using other bacterial species. Owing to specific Salmonella primers, the use of LAMPs approach was time
effective; because these amplicons could be utilized after 20 min instead of the 1 h needed for conventional methods.
Furthermore, LAMP-positive amplicons were rapidly detected within 5 min.
Conclusion, significance and impact study: The determination of DNA in biological samples is a recent keystone in
genomic analysis techniques. Salmonella is a foodborne pathogen that causes many diseases and, in extreme cases,
death. Accordingly, detecting Salmonella has become a vital issue for food safety and security. Combining DNA and
MBs on paper helped us to develop a new method for label-free, non-immobilized, naked eye detection of Salmonella.
The process is very specific owing to the use of exact primers and does not require heavy or expensive instrumentation.
In the future, this method could be applied to biosensors as well as in biomedical and molecular diagnostic fields.
- Full text:P020170105559344339075.pdf
