Table 1.
qPCR primers and probes with target gene, oligonucleotide primer sequence, amplicon size, positive control for generation of standards, and primer/probe reference. Standard curves of 101 to 108 gene copies were used for quantification.
| Target Gene | Primer/ Probe |
Sequence (5′→3′) | Amplicon Size [bp] | Positive Controls | Reference |
|---|---|---|---|---|---|
| blaCTX-M | q_CTXM-F | CAGCTGGGAGACGAAACGTT | 63 | E. coli 1058_16 * | [64] |
| q_CTXM-R | CCGGAATGGCGGTGTTTA | ||||
| q_CTXM-P | 6FAM- CGTCTCGACCGTACCGAGCCGAC-TAMRA |
||||
| blaIMP | q_blaIMP-F | ATTTTCATAGTGACAGCACGGGC | 105 | Klebsiella pneumoniae R77 IMP-4 | [65] |
| q_blaIMP-R | CCTTACCGTCTTTTTTAAGCAGCTCATTAG | ||||
| q_blaIMP-P | 6FAM- TTCTCAACTCATCCCCACGTATGC-TAMRA |
||||
| blaTEM | q_TEM-F | CACTATTCTCAGAATGACTTGGT | 85 | E. coli 832_16_2 * | [66] |
| q_TEM-R | TGCATAATTCTCTTACTGTCATG | ||||
| q_TEM-P | 6FAM- CCAGTCACAGAAAAGCATCTTACGG-TAMRA |
||||
| intI1 | q_intI-F | GCCTTGATGTTACCCGAGAG | 196 | E. coli pKJK5 #337 | [67] |
| q_intI-R | GATCGGTCGAATGCGTGT | ||||
| q_intI-P | 6FAM-ATTCCTGGCCGTGGTTCTGGGTTTT-TAMRA | ||||
| korB | q_korB-F | TCATCGACAACGACTACAACG | 118 |
E. coli RP4 (IncP-1α) E. coli JM109 pB10 (IncP-1ß) E. coli JM109 pQKH545 (IncP-1γ) E. coli JM109 pKJK5 (IncP-1ε) |
[58] |
| q_korB-Fz | TCGTGGATAACGACTACAACG | ||||
| q_korB-R | TTCTTCTTGCCCTTCGCCAG | ||||
| q_korB-Rge | TTYTTCYTGCCCTTGGCCAG | ||||
| q_korB-Rd | TTCTTGACTCCCTTCGCCAG | ||||
| q_korB-P | 6FAM- TCAGYTCRTTGCGYTGCAGGTTCTCVAT-TAMRA |
||||
| q_korB-Pgz | 6FAM- TSAGGTCGTTGCGTTGCAGGTTYTCAAT-TAMRA |
* clinical isolates from Robert Koch Institute Wernigerode.