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Table 1 The oligonucleotide primers and the PCR programs used for amplification of O-serogroups, virulence factors and antibiotic resistance genes of Escherichia coli isolates of hospital foods

From: Shiga (Vero)-toxin producing Escherichia coli isolated from the hospital foods; virulence factors, o-serogroups and antimicrobial resistance properties

Target gene

Primer sequence (5’–3’)

PCR product (bp)

PCR programs

PCR Volume (50 μL)

O157

F: CGGACATCCATGTGATATGG

R: TTGCCTATGTACAGCTAATCC

259

1 cycle:

95 0C ------------ 3 min.

30 cycle:

95 0C ------------ 20 s

58 0C ------------ 40 s

72 0C ------------ 30 s

1 cycle:

72 0C ------------ 8 min

5 μL PCR buffer 10×

2 mM Mgcl2

150 μM dNTP (Fermentas)

0.75 μM of each primers F & R

1.5 U Taq DNA polymerase (Fermentas)

3 μL DNA template

O145

F: CCATCAACAGATTTAGGAGTG

R: TTTCTACCGCGAATCTATC

609

O103

F: TTGGAGCGTTAACTGGACCT

R: GCTCCCGAGCACGTATAAG

321

O26

F: CAGAATGGTTATGCTACTGT

R: CTTACATTTGTTTTCGGCATC

423

O111

F: TAGAGAAATTATCAAGTTAGTTCC

R: ATAGTTATGAACATCTTGTTTAGC

406

O91

F: GCTGACCTTCATGATCTGTTGA

R: TAATTTAACCCGTAGAATCGCTGC

291

1 cycle:

94 0C ------------ 6 min.

34 cycle:

95 0C ------------ 50 s

58 0C ------------ 70 s

72 0C ------------ 55 s

1 cycle:

72 0C ------------ 10 min

5 μL PCR buffer 10×

2 mM Mgcl2

150 μM dNTP (Fermentas)

0.75 μM of each primers F & R

1.5 U Taq DNA polymerase (Fermentas)

3 μL DNA template

O128

F: GCTTTCTGCCGATATTTGGC

R: CCGACGGACTGATGCCGGTGATT

289

O121

F: TGGCTAGTGGCATTCTGATG

R: TGATACTTTAGCCGCCCTTG

322

O113

F: GGGTTAGATGGAGCGCTATTGAGA

R: AGGTCACCCTCTGAATTATGGCAG

771

O45

F: CCGGGTTTCGATTTGTGAAGGTTG

R: CACAACAGCCACTACTAGGCAGAA

527

stx1

F: AAATCGCCATTCGTTGACTACTTCT

R: TGCCATTCTGGCAACTCGCGATGCA

366

1 cycle:

95 0C ------------ 3 min.

34 cycle:

94 0C ------------ 60 s

56 0C ------------ 45 s

72 0C ------------ 60 s

1 cycle:

72 0C ------------ 10 min

5 μL PCR buffer 10×

2 mM Mgcl2

150 μM dNTP (Fermentas)

0.75 μM of each primers F & R

1.5 U Taq DNA polymerase (Fermentas)

3 μL DNA template

stx2

F: CGATCGTCACTCACTGGTTTCATCA

R: GGATATTCTCCCCACTCTGACACC

282

eaeA

F: TGCGGCACAACAGGCGGCGA

R: CGGTCGCCGCACCAGGATTC

629

ehly

F: CAATGCAGATGCAGATACCG

R: CAGAGATGTCGTTGCAGCAG

432

aadA1

F: TATCCAGCTAAGCGCGAACT

R: ATTTGCCGACTACCTTGGTC

447

1 cycle:

94 0C ------------ 8 min.

32 cycle:

95 0C ------------ 60 s

55 0C ------------ 70 s

72 0C ------------ 2 min

1 cycle:

72 0C ------------ 8 min

5 μL PCR buffer 10×

2 mM Mgcl2

150 μM dNTP (Fermentas)

0.75 μM of each primers F & R

1.5 U Taq DNA polymerase (Fermentas)

3 μL DNA template

tetA

F: GGTTCACTCGAACGACGTCA

R: CTGTCCGACAAGTTGCATGA

577

tetB

F: CCTCAGCTTCTCAACGCGTG

R: GCACCTTGCTGATGACTCTT

634

dfrA1

F: GGAGTGCCAAAGGTGAACAGC

R: GAGGCGAAGTCTTGGGTAAAAAC

367

qnr

F: GGGTATGGATATTATTGATAAAG

R: CTAATCCGGCAGCACTATTTA

670

aac (3)-IV

F: CTTCAGGATGGCAAGTTGGT

R: TCATCTCGTTCTCCGCTCAT

286

sul1

F: TTCGGCATTCTGAATCTCAC

R: ATGATCTAACCCTCGGTCTC

822

blaSHV

F: TCGCCTGTGTATTATCTCCC

R: CGCAGATAAATCACCACAATG

768

CITM

F: TGGCCAGAACTGACAGGCAAA

R: TTTCTCCTGAACGTGGCTGGC

462

cat1

F: AGTTGCTCAATGTACCTATAACC

R: TTGTAATTCATTAAGCATTCTGCC

547

cmlA

F: CCGCCACGGTGTTGTTGTTATC

R: CACCTTGCCTGCCCATCATTAG

698