TABLE 10.

List of primers used and thermocycler setting^a used for the amplification of the virulence genes

Set	Target gene	Accession no.	Primer name	Sequence (5′ to 3′)	Length (nt)d	Temp (°C)	Amplicon size (bp)
Ab	ciaB	NP_282066	ciaB.F335	GGTCTAACTTCATCAACCCTTTGC	24	62.9	658
			ciaB.R992	CTCATGCGGTGGCATTAGAATG	22	62.7
	cadF	NP_282616	cadF.F20	GCATCCACTCTTCTATTATCCGC	23	62.8	543
			cadF.R562	ATTCCGTCTTAGTGATTCTTTGGC	24	61.2
	cdtA	NP_281292	cdtA.F3	ATCGTACCTCTCCTTGGCG	19	62.3	440
			cdtAR442	CGGAGCAGCTTTAACGGTTTG	21	62.6
	cdtC	NP_281290	cdtC.F260	GCTCCAAAGGTTCCATCTTCTAAG	24	62.9	263
			cdtC.R522	GCAACTCCTACTGGAGATTTGAAAG	25	62.9
	cdtB	NP_281291	cdtB.F152	GCTTGAGTTGCGCTAGTTGG	20	62.4	180
			cdtB.R331	TGGAGGAACAGATGTAGGAGC	21	62.6
Bb	virB9	YP_980061	virB9.F429	AAGAACACGCTTTGCAATGGC	21	60.6	535
			virB9.R964	CGATGATCCTAGTCTCTACTGGAC	24	64.6
	pebA	NP_282073	pebA.F40	GCTCTAGGTGCTTGTGTTGC	20	62.4	436
			pebA.R476	GTAGTTGCAGCTTGAGCCAC	20	62.4
	porA	NP_282406	porA.F740	TCAACTGGACACTTGAAGGTGC	22	62.7	342
			porA.R1082	CCACCATATACGAAGTCAGCACC	23	64.8
	flhB	NP_281526	flhB.F531	GGTTGCACAGCTTACTTGGC	20	62.4	257
			flhB.R788	ACATCCGCACCTGCAACATC	20	62.4
	jlpA	NP_282133	jlpA.F998	GCACACAGGGAATCGACAGC	20	64.5	119
			jlpA.R1116	AAATGACGCTCCGCCCATTAAC	22	62.7
Cc	flaAB	NP_282485 (flaA), NP_282484 (flaB)	flaA.R1094	CAGTTGGAACAGGACTTGGAG	21	62.6	~1,500
			flaB.R253	GCTCATCCATAGCCTTATCAGCAG	24	64.6
	pldA	Part of NC_002163	pldA.F422	GCCTATACTCAAACTTCTTGGTGG	24	60.6	499
			pldA.R940	AGTCTATAAGGCTTTCTCCATAGCC	25	62.9
	flgB	NP_281712	flgB.F25	GAACTGGTCACTGGTGCTTTAGC	23	64.6	224
			flgB.R248	CTAGGATCAGGGAATTTCCAAGG	23	62.8

^aPCR thermocycler condition was initial denaturation at 95°C for 3 min followed by 35 cycles of 95°C for 30 s, 60°C for 30 s, 72°C for 90 s, and a final extension step of 72°C for 5 min.

^bAntibiotics resistance genes run as a pentaplex PCR.

^cAntibiotics resistance gene run as a triplex PCR.

^dnt, nucleotide.