Table 1 Oligonucleotide sequences of capture-oligonucleotides, real-time PCR primers and probes
No. | Name | Sequence 5′-3′ | 5′-label | 3′-label | |
|---|---|---|---|---|---|
1 | oligonucleotides for capturing MTC DNA: Cap 1 and Cap 2 are specific for Heli; Cap 3 and Cap 4 are specific for IS 1081 | Cap 1 | TTG ATC AGG TCG ACG ATG TAG | Biotin TEG | |
Cap 2 | TCA CCA CCG ACA AAG CGT C | Biotin TEG | |||
Cap 3 | CTC TCG ACG TTC ATC GCC G | Biotin TEG | |||
Cap 4 | TGG CGG TAG CCG TTG CGC | Biotin TEG | |||
2 | duplex real-time PCR specific for the hypothetical Helicase (Heli) | MTC-Heli 4 F | TTG ATC AGG TCG ACG ATG TAG | ||
MTC Heli 4R | TCA CCA CCG ACA AAG CGT C | ||||
MTC Heli 4 FAM | TCA ACG ACC CCA ACG ACT GGT GC | FAM | BHQ1 | ||
3 | duplex real-time PCR specific for the insertion sequence 1081 (IS 1081) | MTC IS 1081 5 F | CTC TCG ACG TTC ATC GCC G | ||
MTC IS 1081 5R | TGG CGG TAG CCG TTG CGC | ||||
MTC IS 1081 5 FAM | ATT GGA CCG CTC ATC GCT GCG TTC | FAM | BHQ1 | ||
4 | Duplex real-time PCR specific for ß-actin | ACT2 1030 F | AGC GCA AGT ACT CCG TGT G | ||
ACT 1135 R | CGG ACT CAT CGT ACT CCT GCT T | ||||
ACT-1081 1105 Y | TCG CTG TCC ACC TTC CAG CAG ATGT | YakimaYellow | BHQ1 |