Table 1. Oligonucleotide primers and probes for the real time PCR to detect target pathogens and an internal control.

Pathogen	Target gene	Type	Sequences (5’-3’)	Tm (°C)	Product size (bp)	Reference
K. pneumoniae	yphG	F. primer	GAGTTAGGGAAACGAACATTGTGIIIIIGGCAGTGCTC	58	137	[16]
		R. primer	TCTCTATCGGACAGACGTCGGIIIIIAAAGAGGTTC	59
		probe	aFAM-TTCATTGGCATCATCACTTAGCGAC	65.1
P. aeruginosa	regA	F. primer	GCTTCATCGACAGCATCGGIIIIIACTGACGCCA	58.1	111	[17]
		R. primer	CGGCTTTTTCTCTGGTCTGTGIIIIIGTTCCGCTGT	59.1
		probe	bHEX-AACACAAACGCACTCGGAAAAATCG	68.5
S. aureus	nuc	F. primer	GATGGCTATCAGTAATGTTTCGAIIIIICAATACRCAA	56.3	213	[18]
		R. primer	GTCGCAGGTTCTTTATGTAATTTTIIIIITGAAGTTGCA	57.4
		probe	cTexas red-CAAGTCTAAGTAGCTCAGCARATGCATCA	67.1
M. catarrhalis	copB	F. primer	ATTCGTGGCATGGGTCATAAT	58.5	182	[9]
		R. primer	GTAACAATCGCACCRTTGGTT	56.8~59.3
		probe	dCy5.5-CACCAAGGTCGCTTTATGCTAGACCC	68.7
Internal control	HBB	F. primer	GGCATAAAAGTCAGGGCAGAIIIIICTATTGCT	56.9	158	-
		R. primer	CCAACTTCATCCACGTTCACCIIIIICCACAGGG	59.0
		probe	eCy5-CCTGAGGAGAAGTCTGCCGTTACTGC	68.8

Probes were labeled with FAM, HEX, Texas red, Cy5.5, and Cy5 and detected at 520, 556, 616, 694, and 669 nm, respectively. All the probes were had BHQ as a quencher at 3’ end.

Abbreviations: T_m, melting temperature; F. primer, forward primer; R. primer, reverse primer; probe, fluorescently labeled primer; R: A or G.