Same-Day Detection of Viable Salmonella Enteritidis by Phage Amplification Coupled with Loop-Mediated Isothermal Amplification
2025
Lamas, Alexandre | Garrido-Maestu, Alejandro
9 pages, 3 tables, 1 figure
Show more [+] Less [-]In recent years, isothermal techniques such as loop-mediated isothermal amplification (LAMP) have appeared as alternatives to qPCR. Due to its amplification at constant temperature, LAMP reactions can be performed in equipment as simple as a thermoblock. However, one of the main limitations of nucleic acid-based techniques is their incapacity to differentiate between live and death cells. To overcome this limitation, some researchers are evaluating the detection of bacteriophages (phages) DNA instead of the detection of bacterial nucleic acids. Phages are used in the sample enrichment step where they reproduce in the presence of live cells. Due to narrow spectrum activity, one phage can infect one bacterial species or even serotype, which allows to perform a selective enrichment. This chapter describes a protocol for the detection of Salmonella Enteritidis in chicken meat in less than 8 h. This method combines sample enrichment in peptone water to which phage PVP-SE2 is added with phage DNA detection by LAMP. In addition, a colorimetric variant of this technique is described that allows this technique to be applied at points of interest
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