Microbial strains Tm P (°C) Tm A (°C) Gram positive (G+) mean SD mean SD Enterococcus faecalis 67.94 0.07 84.14 0.36 Enterococcus faecium 67.84 0.21 84.59 0.78 Listeria monocytogenes 67.80 0.19 86.01 0.36 Staphylococcus aureus 64.85 0.21 83.91 0.54 Staphylococcus epidermidis 64.50 0.30 83.60 0.36 Streptococcus pyogenes 46.54 0.56 84.38 0.78 Gram negative (G-)         Acinetobacter baumannii 66.09 0.15 82.90 0.16 Bacteroides fragilis 48.65 0.18 84.47 0.84 Enterobacter aerogenes 63.95 0.34 83.47 0.48 Enterobacter cloacae 64.98 0.09 84.38 0.24 Escherichia coli 64.69 0.44 84.74 0.54 Haemophilus influenzae 61.99 0.35 84.28 0.30 Klebsiella pneumoniae 65.13 0.23 84.57 0.20 Proteus vulgaris 64.58 0.18 82.87 0.24 Pseudomonas aeruginosa 53.32 0.33 83.00 0.34 Serratia marcescens 64.01 0.30 84.17 0.30 Stenotrophomonas maltophilia 58.10 0.07 84.42 0.15 Fungi         Candida albicans - - 87.1 0.33 Candida dubliniensis - - 85.5 0.50 Candida quillermondii - - 85.1 0.70 Candida krusei - - 89.8 0.02 Candida parapsilosis - - 85.4 0.88 Candida tropicalis - - 84.5 0.75 Aspergillus fumigatus - - 91.0 0.38

Table 1: Melting points of bacterial and fungal amplicons and probes. All the ampli- cons Tm measured at the F1 channel (540 nm). The signal generated by a specific SYBR Green dye. The G+ specific probes got the signal at the F2 channel (640 nm) the G- probes at the F3 channel (705 nm). The signal induced by the help of special FRET technique.