Table 2.

The oligonucleotide primers and the Multiplex PCR programs used for amplification of virulence genes of E. coli isolates

Gene	Primer name	Primer sequence (5'-3)	Size of product (bp)	PCR program	M-PCR volume (50 μL)
pap	pap3 pap4	GCAACAGCAACGCTGGTTGCATCAT AGAGAGAGCCACTCTTATACGGACA	336	1 cycle: 94 °C ------------ 1 min. 30 cycle: 94 °C ------------ 60 s 63 °C ------------ 30 s 72 °C ------------ 90 s 1 cycle: 72 °C ------------ 5 min	5 μL PCR buffer 10X 1.25 mM Mgcl2 150 μM dNTP (Fermentas) 1 μM of each primers F & R 1.2 U Taq DNA polymerase (Fermentas) 3 μL DNA template
Sfa	sfa1 sfa2	CTCCGGAGAACTGGGTGCATCTTAC CGGAGGAGTAATTACAAACCTGGCA	410
Afa	afa1 afa2	GCTGGGCAGCAAACTGATAACTCTC CATCAAGCTGTTTGTTCGTCCGCCG	750
fimH	FimH1 FimH2	GAGAAGAGGTTTGATTTAACTTATTG AGAGCCGCTGTAGAACTGAGG	559	1 cycle: 94 °C ------------ 3 min. 40 cycle: 94 °C ------------ 60 s 58 °C ------------ 70 s 72 °C ------------ 70 s 1 cycle: 72 °C ------------ 6 min	5 μL PCR buffer 10X 2 mM Mgcl2 200 μM dNTP (Fermentas) 0.4 μM of each primers F & R 1 U Taq DNA polymerase (Fermentas) 3 μL DNA template