Table 1 Primers and probes for multiplex TaqMan PCR
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 |