Table 1.
Primers DNA sequences used in the study
| Primer sequence (5'- 3') | Target | PCR program | Size of amplicon (bp) |
|---|---|---|---|
| First multiplex PCR | |||
| GGGGTTCTGAGAGGAAG ACCCCCCATTGTACCAC |
Bacteroides spp.9 | 4 min at 94°C; 30 cycles (45 sec at 94°C, 45 sec at 60°C, and 1 min at 72°C); 7 min at 72°C (this study) | 950 |
| CTCAACTTGGGTGCTGCATTT ATTGTAGTACGTGTGTAGCCC |
Clostridioides spp.9 | 619 | |
| GGTGCCGCAGTAAACACAATAAGT AAGGCCCGGGAACGTATTCA |
Peptococcus spp.9 | 539 | |
| CCTCTGACCGCTCTTTAATCGGAGCTTTCCTTC CCAGTTATCGGTCCCACCTTCGGCAGCT |
Ruminococcus spp.9 | 482 | |
| AACTCCGGTGGTATCAGATG GGGGCTTCTGAGTCAGGTA |
Peptostreptococcus spp.9 | 270 | |
| Second multiplex PCR | |||
| GTTTGGTCGCATATCGCAAC AATGCGCAGCACCAGGATAG |
bla VIM variants10 | 4 min at 94°C; 30 cycles (45 sec at 94°C, 45 sec at 55°C, and 1 min at 72°C); 7 min at 72°C (this study) | 382 |
| GAAGGYGTTTATGTTCATAC GTAMGTTTCAAGAGTGATGC |
bla IMP variants10 | 587 | |
| GGGCAGTCGCTTCCAACGGT GTAGTGCTCAGTGTCGGCAT |
bla NDM-1 10 | 475 | |
| TCTCGGGTAACATCAAGG AGGAGATCCGAAGACCTC |
intI-1 11 | 234 | |
| Third multiplex PCR12 | |||
| GCAGCTTGTCGGCCATGCGGGC GGTCGCGAAGCTGAGCACCGCAT |
bla NDM-1 | 5 min at 95°C; 35 cycles (45 sec at 95°C, 45 sec at 60°C, and 1 min at 72°C); 8 min at 72°C | 782 |
| GCGTGGTTAAGGATGAACAC CATCAAGTTCAACCCAACCG |
bla OXA-48 | 438 | |
| TGTCACTGTATCGCCGTC CTCAGTGCTCTACAGAAAACC |
bla KPC | 900 | |
| GAAGGCGTTTATGTTCATAC GTACGTTTCAAGAGTGATGC |
bla IMP | 587 | |
| GTTTGGTCGCATATCGCAAC AATGCGCAGCACCAGGATAG |
bla VIM-1 | 389 | |
| Internal control genes | |||
| CAACTTCATCCACGTTCACC GAAGAGCCAAGGACAGGTAC |
Human β-globin13 | 268 | |
| AGTTTGATCCTGGCTCAG GGACTACCAGGGTATCTAAT |
Bacterial 16SrRNA13 | 798 | |