Molecular Characterization and Antimicrobial Resistance of Pathogenic Escherichia coli Strains in Children from Wolaita Sodo, Southern Ethiopia

Table 1

Primer sequences specific for each virulence gene, their amplicon size, and optimized PCR protocol.

Target gene	Primer name	Sequence 5′ to 3′	Amplicon size (bp)	PCR amplification						Escherichia coli strain detected	References
				Initial denaturation  C/min	Denaturation °C/sec.	Annealing °C/sec	Extension °C/sec	Final extension °C/min	No. of cycle
eaeA	EAE1	F: AAACAGGTGAAACTGTTGCC	490	95/3	95/40	55/60	72/60	72/10	35	EPEC/EHEC	[44]
	EAE2	R: CTCTGCAGATTAACCTCTGC
Stx 1	EVS1	F: ATCAGTCGTCACTCACTGGT	110	95/3	95/40	57/40	72/30	72/8	30	STEC/EHEC	[45]
	EVC2	R: CTGCTGTCACAGTGACAAA
Stx 2	EVT1	F: CAACACTGGATGATCTCAG	350	95/3	95/40	57/40	72/30	72/8	30	STEC/EHEC
	EVT2	R: CCCCCTCAACTGCTAATA
Bfp	bfpA1	F: TTCTTGGTGCTTGCGTGTCTTTT	324	95/3	95/40	57/40	72/30	72/8	30	Typical EPEC	[44]
	bfpA2	R: TTTTGTTTGTTGTATCTTTGTAA
HlyA	hlyA-1	F: GGT GCA GCA GAA AAA GTT GTA G	190	95/3	95/40	45/60	72/60	72/10	30	EHEC	[46]
	hlyA-2	R: TCT CGC CTG ATA GTG TTT GGT a
aatA	aatA1	F: GGT GCA GAA CAC GAT GCA	590	95/3	95/40	45/60	72/60	72/10	30	EAEC	[44]
	aatA2	R: CCA CGT CTT GTG CTA CGT
Lt	LT-1	F: TCTCTATGTGCATACGGAGC	696	95/3	95/40	53/60	72/30	72/8	30	ETEC	[47]
	LT-2	R: CCATACTGATTGCCGCAAT
St	ST-1	F: TGCTAAACCAGTAGAGTCTTCAAAA	294	95/3	95/40	52/60	72/60	72/10	35	ETEC	[48]
	ST-2	R:GCAGGATTACAACACAATTCACAGCAG
Dae	daeE 1	F: GAA CGT TGG TTA ATG TGG GGT AA	542	95/3	95/40	45/60	72/60	72/10	37	DAEC	[49]
	daeE 2	R: TAT TCA CCG GTC GGT TAT CAG

EPEC = enteropathogenic Escherichia coli; ETEC = enterotoxigenic Escherichia coli; STEC = shiga toxin-producing Escherichia coli; EHEC = enterohaemorrhagic Escherichia coli; EAEC = enteroaggregative Escherichia coli; DAEC = diffusely adherent Escherichia coli; EAE = effacing and attaching, Stx = shiga toxin, bp = Base pair.