Table 5 Primer sequences used for RT-PCR to identify the nine target genes for seven foodborne bacteria from the extracted DNA samples.

Bacteria	Gene	Sequences (5ʹ–3ʹ)	Accession number	Amplicon size	Primer concentration^a	R^2a	Efficiency (%)^a
Staphylococcus aureus	htrA	GAAGTAATATCAGACA AATCAAATCAGTACC	NC_009782	92 bp	500 nM	0.996	86.2
Staphylococcus aureus	htrA	TCTTCCGGTAAAGTTAATGGCTTCTG	NC_009782	92 bp	500 nM	0.996	86.2
Escherichia coli	yccT	GCAGCGTGGTGGCAAAA	CP10315	56 bp	400 nM	1.00	91.6
Escherichia coli	yccT	CGTGACCACCTTGATTGCAT	CP10315	56 bp	400 nM	1.00	91.6
STEC Escherichia coli	eae	GTAACAATGTCAGAGG CGAGTTG	AE005174	72 bp	600 nM	0.998	92.1
	eae	CCACCGCTTGCTTTCAGTTTAA	AE005174	72 bp	600 nM	0.998	92.1
	stx1	GGATTTCGTACAACACTGGATGATC	M16625	67 bp	400 nM	0.998	89.9
	stx1	GATCAACATCTTCAGCA GTCATTACA	M16625	67 bp	400 nM	0.998	89.9
	stx2	ACCCCACCGGGCAGTT	X07865	59 bp	500 nM	0.999	98.3
	stx2	CGCGCCTGATAGACATCAAG	X07865	59 bp	500 nM	0.999	98.3
Listeria monocytogenes	inlA	TAACAGACACGGTCTCG CAAA	CP013288	66 bp	400 nM	0.999	93.9
Listeria monocytogenes	inlA	TCCCTAATCTATCCGCCTGAAG	CP013288	66 bp	400 nM	0.999	93.9
Yersinia enterocollitica	yst	TGGAGCATTCGGCCAA GAA	X65999	60 bp	400 nM	0.995	92.2
Yersinia enterocollitica	yst	ATTGGTGTCGATAATG CATCACTGA	X65999	60 bp	400 nM	0.995	92.2
Salmonella enterica	invA	TGGAAAGGGAAAGCC AGCTT	M90846	68 bp	500 nM	0.995	87.8
Salmonella enterica	invA	AATAGCGTCACCTTTG ATAAACTTCA	M90846	68 bp	500 nM	0.995	87.8
Campylobacter jejuni	cadF	TGAACCAAGAGAAGG TGCTTTGT	FJ946061	76 bp	400 nM	0.998	97.5
Campylobacter jejuni	cadF	AAAACCAAAATGACCTTCCAAAGAAATAGTT	FJ946061	76 bp	400 nM	0.998	97.5

^aPrimers were selected from previously published studies^77,78. The appropriate primer concentration was selected based on the reproducibility (R² > 0.99) and efficiency (> 85%) of the selected primers (forward and reverse) to detect the targeted genes specific to each of the ATCC positive controls (S. aureus ATCC 19048, E. coli ATCC 11229, E. coli O157:H7 ATCC 35150, Listeria monocytogenes ATCC 13932, Yersinia enterocolitica DSM 4780, Salmonella enterica ATCC 13076, and Campylobacter jejuni ATCC 33291). To calculate R² and efficiency (%), the chosen primers for the targeted genes specific to each ATCC strain were run with duplicates in fourfold serial dilutions (undiluted, 1:3, 1:9, 1:27, 1:81); a negative control was also run (Nuclease free water).

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