Bacterial Detection in Milk Samples

The complete method (DED–MIRA–LFD) consisted of a DED for extraction and MIRA–LFD for amplification and detection. To determine the specificity and sensitivity of DED–MIRA–LFD with the selected primer, 10 μl of the bacterial solution was added to 90 μl of sterile milk as artificially spiked milk. Specificity was conducted with 12 species of bacteria-spiked milk including S. agalactiae; the negative control included a sterile milk sample instead of a spiked milk sample. Sensitivity was validated using a 10-fold serially diluted bacterial solution of spiked milk ranging from 10¹ to 10⁷ CFU/ml.