| Full text | |
| Author(s): |
Serain, Alessandra Freitas
;
deSouza, Daniel Rangel
;
Viviani, Vadim R.
Total Authors: 3
|
| Document type: | Journal article |
| Source: | CHEMOSENSORS; v. 13, n. 2, p. 14-pg., 2025-02-01. |
| Abstract | |
New faster and more sensitive detection methods are required for food microbiological contamination quality control throughout the entire handling process. The adenosine triphosphate (ATP) bioluminescence detection method based on the firefly luciferin-luciferase system has been a widely used approach for decades due to its practicality, efficiency, and rapidity. The luciferase of the Amydetes vivianii firefly, cloned and produced in our laboratory, displays a high bioluminescence and stability, showing desirable properties for ATP assays. Using this enzyme, in this study we developed and validated a swab-based ATP detection method for meat, chicken, and milk, allowing us to distinguish between contaminated and non-contaminated forms of these foods. This method demonstrated robust interday and intraday precision, an accuracy of 70-71% across different matrices, and a limit of detection of 1.0 x 10(4) cps for the dilutions and 2.7 x 10(3) cps for the swabs, fulfilling all validation criteria and ensuring reliability for routine applications. Except for milk, which has very low endogenous ATP levels due to pasteurization, thus requiring sample pre-processing, the method allowed us to luminometrically detect ATP on the surface of meat and chicken in less than an hour using an assay solution. The method showed higher sensitivity compared to an available commercial kit due to its intense signal, with a remarkable similar to 22-fold increase in luminescence intensity when comparing the highest ATP concentration of Amydetes luciferase with a commercially available luciferase, allowing for detecting microbiological contamination at lower levels or using less sensitive luminometers. (AU) | |
| FAPESP's process: | 10/05426-8 - Arthropod bioluminescence: biological diversity in Brazilian biomes, biochemical origin, structural/functional evolution of luciferases, molecular differentiation of lanterns, biotechnological, environmental and educational applications |
| Grantee: | Vadim Viviani |
| Support Opportunities: | Research Projects - Thematic Grants |
| FAPESP's process: | 21/03901-5 - Luciferases expression in mammalian cells for bioassay purposes and phototoxic effect of luciferins and fluorescent compounds from coleoptera and diptera |
| Grantee: | Alessandra Freitas Serain |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 17/22262-8 - Multiusuary equipment conceeded by process: 2010/05426-8-bioimaging CCD system |
| Grantee: | Vadim Viviani |
| Support Opportunities: | Multi-user Equipment Program |
| FAPESP's process: | 18/17855-2 - Cloning, expression and characterization of Phengodidae luciferases and proteins of bioluminescent system: structural, functional and evolutive aspects |
| Grantee: | Daniel Rangel de Souza |
| Support Opportunities: | Scholarships in Brazil - Doctorate (Direct) |