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Table 1 Primers and probes for multiplex TaqMan PCR

From: A set of novel multiplex Taqman real-time PCRs for the detection of diarrhoeagenic Escherichia coli and its use in determining the prevalence of EPEC and EAEC in a university hospital

Target Primer or probe name Oligonucleotide sequence (5'→3') Amplicon size/Accession nr.
Multiplex 1: (EPEC/EAEC)   
EAF EP-1 for
EP-2 rev
EP-S probe
GTT CTT GGC GAA CAG GCT TGT C
TTA AGC CAG CTA CCA TCC ACC C
Cy5-AGT ACT GAC GTG CAG GTC GCC TGT TCG-BHQ-3
107 bp
X76137
eae EAE-S for
EAE-B1 rev
EAE-B2 rev
EAE-TM probe
ACT GGA CTT CTT ATT RCC GTT CTA TG
CTA AGC GGG TAT TGT TAC CAG A
CCT AAA CGG GTA TTA TCA CCA GA
ROX-AAT CCT GAT CAA TGA AGA CGT TAT AGC CCA-BBQ
189 bp*
Z11541/
AB040740
pCVD432 EA-1 for
EA-2 rev
EA-S probe
AGG TTT GAT ATT GAT GTC CTT GAG GA
TCA GCT AAT AAT GTA TAG AAA TCC GCT GTT
FAM-CAT GTT CCT GAG AGT GCA ATC CCA GAC ATT AC-TAMRA
152 bp
X81423
Multiplex 2: (ETEC/EIEC)   
ST gene ST-1 for
ST-2 rev
ST-S probe
CTG GTT TTG ATT CAA ATG TTC GTG
TCC TGA GGG AAA GGT GAA AAA GAC
ROX-TTG ATT TCT TCA TAT TAC CTC CGG ACA TGG CA-BHQ-2
107 bp
M34916
LT gene LT-1 for
LT-2 rev
LT-S probe
AGC GGC GCA ACA TTT CAG
TTG GTC TCG GTC AGA TAT GTG ATT C
FAM-TCG AAG TCC CGG GCA GTC AAC ATA TAG A-TAMRA
113 bp
S60731
ipa H Ei-1 for
Ei-2 rev
Ei-S probe
GAA CTC AAA TCT TGC ACC ATT CA
CGT CCG TCC GAG AAC AAT TAA G
Cy5-ATC CCC GAC ACC GTT TGT GAG TTT CAC T-BHQ-3
107 bp
AY206439
Multiplex 3: (EHEC)   
stx 1 slt1-1 for
slt1-2 rev
slt1-S probe
CTT CCA TCT GCC GGA CAC ATA
ATT AAT ACT GAA TTG TCA TCA TCA TGC AT
ROX-AAG GAA ACT CAT CAG ATG CCA TTC TGG CA-BHQ-2
87 bp
Z36899
stx 2 slt2-1 for
slt2-2 rev
slt2-S probe
GAC GTG GAC CTC ACT CTG AAC TG
TCC CCA CTC TGA CAC CAT CC
FAM-TAC TCC GGA AGC ACA TTG CTG ATT CGC-TAMRA
82 bp
L11079
Internal Control    
Drosophila simulans + IC probe HEX-ATG CCT CTT CAC ATT GCT CCA CCT TTC CT-BHQ1 AB110070
  1. +The bold, italic labelled primers also detect the internal control plasmid in the respective PCR
  2. *Alignment of various intimin subtype sequences