Table 2 Oligonucleotide primers used in PCR and direct sequencing
Target gene | Primer ID | Nucleotide sequence (5'-3') | Size (bp) | Ref. no. |
|---|---|---|---|---|
MTC identification | ||||
16S rRNA | 285 | GAGAGTTTGATCCTGGCTCAG | 1028 | 7 |
264 | TGCACACAGGCCACAAGGGA | |||
259 | TTTCACGAACAACG GACAA | 591 | ||
cfp32 | Rv0577F | ATGCCCAAGAGAAGCGAATACAGGCAA | 786 | 14 |
Rv0577R | CTATTGCTGCGGTGCGGGCTTCAA | |||
RD9 | Rv2073cF | TCGCCGCTGCCAGATGAGTC | 600 | 14 |
Rv2073cR | TTTGGGAGCCGCCGGTGGTGATGA | |||
RD12 | Rv3120F | GTCGGCGATAGACCATGAGTCCGTCTCCAT | 404 | 14 |
Rv3120R | GCGAAAAGTGGGCGGATGCCAG | |||
RD1 | ET1 | AAGCGGTTGCCGCCGACCGACC | 15 | |
ET2 | CTGGCTATATTCCTGGGCCCGG | |||
ET3 | GAGGCGATCTGGCGGTTTGGGG | |||
RD4 | Rv1510F | GTGCGCTCCACCCAAATAGTTGC | 1033 | 15 |
Rv1510R | TGTCGACCTGGGGCACAAATCAGTC | |||
RD7 | Rv1970F | GCGCAGCTGCCGGATGTCAAC | 1116 | 15 |
Rv1970R | CGCCGGCAGCCTCACGAAATG | |||
MiD3 | IS1561F | GCTGGGTGGGCCCTGGAATACGTGAACTCT | 530 | 15 |
IS1561R | AACTGCTCACCCTGGCCACCACCATGGACT | |||
Distinguish sub-strains of BCG | ||||
RD2 | RD2l | CCAGATTCAAATGTCCGACC | 16 | |
RD2r | GTGTCATAGGTGATTGGCTT | |||
RD14 | RD14l | CAGGGTTGAAGGAATGCGTGTC | 16 | |
RD14r | CTGGTACACCTGGGGAATCTGG | |||
RD15 | RD8l | ACTCCTAGCTTTGCTGTGCGCT | 16 | |
RD8r | GTACTGCGGGATTTGCAGGTTC | |||
RD16 | RD16nf | ACATTGGGAAATCGCTGCTGTTG | 17 | |
RD16nr | GGCTGGTGTTTCGTCACTTC | |||
SenX3-RegX3 | C3 | GCGCGAGAGCCCGAACTGC | 16 | |
C5 | GCGCAGCAGAAACGTCAGC | |||
Sequencing | ||||
mpt64 (Rv1980c) | mpb64W-F | ACTCAGATATCGCGGCAATC | 1061 | this study |
mpb64W-R | CGATCACCTCACCTGGAGTT | |||
Rv1977 | Rv1977F | GTTTCCCGAGATCAGCTCAA | 348 | this study |
Rv1977R | ATCTCGTCGTGTGTCACCAG | |||
Rv1981c | Rv1981F | GATCGAATGCAGGCTGGTAT | 399 | this study |
Rv1981R | ACTACTACCGCGGTGACGAC | |||