Table 2 Primers used.
Primer sequences | Target sequences | References |
---|---|---|
Autotransporters | ||
SPATE1 5′ GAGGTCAACAACCTGAACAAACGTATGGG | The genes encoding SPATE | This study |
SPATE2 5′ CCGGCACGGGCTGTCACTTTCCAG | ||
E coli phylogenetic groups | ||
ChuAf 5′ CGGACGAACCAACGGTCAGGAT | The chuA gene is required for haemtransport in E coli O157:H7 | Modified from Clermont et al15 |
ChuAr 5′ TGCCGCCAGTACCAAAGACACG | ||
Yjaf 5′ CGTGAAGTGTCAGGAGACGCTGC | The yjaA gene coding for protein of unknown function | Modified from Clermont et al15 |
Yjar 5′ TGCGTTCCTCAACCTGTGACAAACC | ||
Tsp1 5′ GGGAGTAATGTCGGGGCATTCAG Tsp2 5′ CATCGCGCCAACAAAGTATTACGCAG | Tsp encodes for a putative DNA fragment (TSPE4.C2) in E coli | Modified from Clermont et al15 |
E coli toxins | ||
Cnff 5′ AGTACTGACACTCACTCAAGCCGC | Cytotoxic necrotising factors (Cnf1 and Cnf2) | This study |
Cnfr 5′ GCAGAACGACGTTCTTCATAAGTATCACC | ||
IpgDf 5′ CGACTTCTCTTCTGACGCCGAC IpgDr 5′ CAACATTCCTCCAGCCTAAGCCC | ipgD gene modulates entry of bacteria into epithelial cells | This study |
Vt1f 5′ CGCATAGTGGAACCTCACTGACGC | Verocytotoxin 1 | This study |
Vt1r 5′ CATCCCCGTACGACTGATCCC | ||
Vt2f 5′ CGGAATGCAAATCAGTCGTCACTCAC | Verocytotoxin 2 | This study |
Vt2r 5′ TCCCCGATACTCCGGAAGCAC | ||
HlyAf 5′ TGCAGCCTCCAGTGCATCCCTC | The hlyA gene encoding α haemolysin | |
HlyAr 5′ CTTACCACTCTGACTGCGATCAGC | ||
STaf 5′ GTGAAACAACATGACGGGAGG | Heat‐stable enterotoxin 1 | This study |
STar 5′ ATAACATCCAGCACAGGCAGG | ||
STbf 5′ GGGGTTAGAGATGGTACTGCTGGAG | Heat‐stable enterotoxin 2 | This study |
STbr 5′ GACAATGTCCGTCTTGCGTTAGGAC | ||
LTf 5′ CCGTGCTGACTCTAGACCCCCA | Heat‐labile enterotoxin LT | This study |
LTr 5′ CCTGCTAATCTGTAACCATCCTCTGC | ||
Eaef 5′ CCAGGCTTCGTCACAGTTGCAGGC Eaer 5′ CGCCAGTATTCGCCACCAATACC | The eae gene coding for intimin present in AEEC strains | This study |
Enterotoxigenic Bacteroides fragilis (ETBF) | ||
Bf1F 5′ GTTAGTGCCCAGATGCAGGATGCGG | The genes coding for Bacteroides fragilis enterotoxins (Bft1, Bft2 and Bft3) | This study |
Bf2F 5′ GAACTCGGTTTATGCAGTTCATGGACTG | ||
Bf3R 5′ TGGGTTGTAGACATCCCACTGGCTT | ||
Bf4R 5′ GGATACATCAGCTGGGTTGTAGACATCCC | ||
E coli adhesins | ||
PapF 5′ CCGGCGTTCAGGCTGTAGCTG PapR 5′ GCTACAGTGGCAGTATGAGTAATGACCGTTA | The genes coding for pathogenicity islands (PAI I, PAI II)16 and sfp gene cluster | This study |
PAI1 5′ TAGCTCAGACGCCAGGATTTTCCCTG | ||
pai2 5′ CCTGGCGCCTGCGGGCTGACTATCAGGG | ||
BmaEf 5′ CTAACTTGCCATGCTGTGACAGTA | The bmaE gene for M‐agglutinin subunit; Afa‐8 gene cluster | Martin et al17 |
BmaEr 5′ TTATCCCCTGCGTAGTTGTGAATC | ||
Sfaf 5′ CGGAGGAGTAATTACAAACCTGGCA | S‐fimbrial adhesins encoded by sfaD to sfaE | Martin et al17 |
Sfar 5′ CTCCGGAGAACTGGGTGCATCTTAC | ||
Afaf 5′ TATGGTGAGTTGGCGGGGATGTACAGTTACA | AfaE‐3 gene cluster | This study |
Afar 5′ CCGGGAAAGTTGTCGGATCCAGTGT | ||
AIDA1 5′ TATGCCACCTGGTATGCCGATGAC AIDA2 5′ ACGCCCACATTCCCCCAGAC | The aidA gene coding E coli AIDA‐I adhesin in DAEC strains | This study |
AggRf 5′ GAGTTAGGTCACTCTAACGCAGAGTTG | The aggR gene for adhesin of aggregative adherence fimbria I | This study |
AggRr 5′ GACCAATTCGGACAACTGCAAGCATCTAC | ||
Ag43F 5′TGACACAGGCAATGGACTATGACCG Ag43R 5′GGCATCATCCCGGACCGTGC | The agn43 gene coding for antigen involved in E coli autoaggregation | This study |
Flagella and type 1 fimbrae typing | ||
Primer1 5′CAAGTCATTAATAC(A/C)AACAGCC | The fliC genes coding for flagellin proteins | Machado et al18 |
Primer2 5′GACAT(A/G)TT(A/G)GA(G/A/C)ACTTC(G/C)GT | ||
FimHf 5′ CTGGTCATTCGCCTGTAAAACCGCCA FimHr 5′ GTCACGCCAATAATCGATTGCACATTCCCT | The fimH gene encoding FimH subunit of type 1 pili | This study |
Ribosomal DNA‐based primers | ||
ITSF 5′ GTCGTAACAAGGTAGCCGTA | 16S‐23S rRNA intergenic transcribed spacers | Cardinale et al13 |
ITSReub 5′ GCCAAGGCATCCACC | ||
27f 5′ AGAGTTTGATCMTGGCTCAG | Conserved 16S rDNA used to amplify ribosomal genes | Lane19 |
342r 5′ CTGCTGCSYCCCGTAG |
SPATE, serine protease autotransporter.