RT Journal Article
T1 Phage amplification coupled with loop-mediated isothermal amplification (PA-LAMP) for same-day detection of viable Salmonella Enteritidis in raw poultry meat
A1 Lamas Freire, Alexandre
A1 Santos, Sílvio
A1 Prado Rodríguez, Marta
A1 Garrido Maestu, Alejandro
K1 Phage amplification
K1 LAMP
K1 Salmonella Enteritidis
K1 Detection
K1 Chicken
K1 Poultry meat
K1 Visual detection
AB Salmonella Enteritidis is the main serotype responsible for human salmonellosis in the European Union. One of the main sources of Salmonella spp. in the food chain are poultry products, such as eggs or chicken meat. In recent years, molecular methods have become an alternative to culture dependent methods for the rapid screening of Salmonella spp. In this work, the strain S. Enteritidis S1400, and previously isolated and characterized bacteriophage PVP-SE2, were used to develop and evaluate a same-day detection method combining Phage Amplification and Loop-mediated isothermal amplification (PA-LAMP) to specifically detect viable S. Enteritidis in chicken breast. This method is based on the detection of the phage DNA rather than bacterial DNA. The virus is added to the sample during pre-enrichment in buffered peptone water, where it replicates in the presence of viable S. Enteritidis. The detection of phage DNA allows, on the one hand to detect viable bacteria, since viruses only replicate in them, and on the other hand to increase the sensitivity of the method since for each infected S. Enteritidis cell, hundreds of new viruses are produced. Two different PA-LAMP detection strategies were evaluated, a real time fluorescence and a naked-eye detection. The present method could down to 0.2 fg/μL of pure phage DNA and a concentration of viral particles of 2.2 log PFU/mL. After a short Salmonella recovery step of 3 h and a co-culture of 4 h of the samples with phage particles, both real-time fluorescence and naked-eye method showed a LoD95 of 6.6 CFU/25 g and a LoD50 of 1.5/25 g in spiked chicken breast samples. The entire detection process, including DNA extraction and LAMP analysis, can be completed in around 8 h. In the current proof-of-concept, the novel PA-LAMP obtained comparable results to those of the reference method ISO 6579, to detect Salmonella Enteritidis in poultry meat.
PB Elsevier
SN 1095-9998
YR 2023
FD 2023-07-10
LK https://hdl.handle.net/10347/43457
UL https://hdl.handle.net/10347/43457
LA eng
NO Food Microbiology Volume 115, October 2023, 104341
NO Dr. Alexandre Lamas was funded by a postdoctoral fellowship from Xunta de Galicia (Axudas de apoio á etapa de formación posdoutoral IN606B (Modalidade A)). Dr. Alejandro Garrido-Maestu acknowledges funding from the Fundação para a Ciência e Tecnologia through the Scientific Employment Stimulus Program (2021.02810.CEECIND).
DS Minerva
RD 24 abr 2026