A valid solution for micro-analytical systems is the selection of a compatible amplification reaction with a
simple, highly-integrated efficient design that allows the detection of multiple genomic targets. Two
approaches under isothermal conditions are presented: recombinase polymerase amplification (RPA)
and multiple displacement amplification (MDA). Both methods were applied to a duplex assay specific
for Salmonella spp. and Cronobacter spp., with excellent amplification yields (0.2–8.6 108 fold). The proposed
approaches were successfully compared to conventional PCR and tested for the milk sample analysis
as a microarray format on a compact disc (support and driver). Satisfactory results were obtained in
terms of resistance to inhibition, selectivity, sensitivity (101
–102 CFU/mL) and reproducibility (below
12.5%). The methods studied are efficient and cost-effective, with a high potential to automate microorganisms
detection by integrated analytical systems working at a constant low temperature.