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Table 1 Oligonucleotides sequences and cycling conditions of PCR assay

From: Virulence determinant and antimicrobial resistance traits of Emerging MDR Shiga toxigenic E. coli in diarrheic dogs

Target genes

Primers sequences

Amplicon size (bp)

Reaction volume (µL)

Amplification (35 cycles)

References

Denat

Annealing

Extension

eaeA (Intimin)

ATGCTTAGTGCTGGTTTAGG

GCCTTCATCATTTCGCTTTC

248

25

94 ˚C

30 s

51 ˚C

30 s

72 ˚C

30 s

(Bisi-Johnson et al. 2011)

hlyA (Hemolysin)

AACAAGGATAAGCACTGTTCTGGCT

ACCATATAAGCGGTCATTCCCGTCA

1177

25

94 ˚C

30 s

54 ˚C

40 s

72 ˚C

45 s

(Piva et al. 2003)

stx1 (Shiga-toxin 1)

ACACTGGATGATCTCAGTGG

CTGAATCCCCCTCCATTATG

614

50

94 ˚C

30 s

58 ˚C

40 s

72 ˚C

45 s

(Dipineto et al. 2006)

stx2 (Shiga-toxin 2)

CCATGACAACGGACAGCAGTT

CCTGTCAACTGAGCAGCACTTTG

779

sul1 (Sulfonamides resistance)

CGGCGTGGGCTACCTGAACG

GCCGATCGCGTGAAGTTCCG

433

25

94 ˚C

30 s

55 ˚C

45 s

72 ˚C

45 s

(Ibekwe et al. 2011)

tetA (Tetracycline resistance)

GGTTCACTCGAACGACGTCA

CTGTCCGACAAGTTGCATGA

576

50

94 ˚C

30 s

55 ˚C

40 s

72 ˚C

45 s

(Randall 2004)

tetB (Tetracycline resistance)

CCTCAGCTTCTCAACGCGTG

GCACCTTGCTCATGACTCTT

634

qnrA (Quinolones resistance)

AGAGGATTTCTCACGCCAGG

TGCCAGGCACAGATCTTGAC

580

50

95 ˚C

1 min

54 ˚C

1 min

72 ˚C

1 min

(Cattoir et al. 2007)

qnrB (Quinolones resistance)

GGMATHGAAATTCGCCACTG

TTTGCYGYYCGCCAGTCGAA

264

qnrS (Quinolones resistance)

GCAAGTTCATTGAACAGGGT

TCTAAACCGTCGAGTTCGGCG

428

blaCTX-M (Cephalosporins resistance)

ATGTGCAGYACCAGTAARGTKATGGC

TGGGTRAARTARGTSACCAGAAYCAGCGG

593

25

94 ˚C

30 s

54 ˚C

40 s

72 ˚C

45 s

(Archambault et al. 2006)

blaTEM (Penicillin resistance)

ATCAGCAATAAACCAGC

CCCCGAAGAACGTTTTC

516

25

94 ˚C

30 s

54 ˚C

40 s

72 ˚C

45 s

(Colom et al. 2003)

blaKPC (Carbpenem resistance)

ATGTCACTGTATCGCCGTCT

TTACTGCCCGTTGACGCCC

892

25

94 ˚C

1 min

55 ˚C

1 min

72 ˚C

1 min

(Xia et al. 2012)

blaNDM-1 (Carbpenem resistance)

GGCGGAATGGCTCATCACGA

CGCAACACAGCCTGACTTTC

287

25

94 ˚C

30 s

55 ˚C

30 s

72 ˚C

30 s