TABLE 2.
Primers used for the MAPCR procedure
| Primer | Target(s) | Sequence (5′-3′)a | Concn(s) (nM) | Tm(s) (°C)b |
|---|---|---|---|---|
| Mox-uf | blaMOX-1, blaCMY-1, and blaCMY-8 | Uni-CTGCTCAAGGAGCACAGGAT | 200 | 71.9 |
| Mox-r to blaCMY-11 | CACATTGACATAGGTGTGGTGC | 200/40/20/10 | 58.3/56.3/55.4/54.5 | |
| cit-uf | blaLAT-1 to blaLAT-4, and blaCMY-2 to blaCMY-7 | Uni-TGGCCAGAACTGACAGGC | 200 | 72.1 |
| cit-r | TTTCTCCTGAACGTGGCTGGC | 200/40/20/10 | 61.7/59.6/58.7/57.8 | |
| dha-uf | blaDHA-1 and blaDHA-2 | Uni-AACTTTCACAGGTGTGCTGGGT | 100 | 71.3 |
| dha-r | CCGTACGCATACTGGCTTTGC | 100/20/10/5 | 60.0/57.9/57.0/56.1 | |
| acc-uf | blaACC-1 and blaACC-2 | Uni-AACAGCCTCAGCAGCCGGTTA | 100 | 72.4 |
| acc-r | TTCGCCGCAATCATCCCTAGC | 100/20/10/5 | 60.8/58.7/57.8/56.9 | |
| ebc-uf | blaMIR-1 and blaACT-1 | Uni-CAGGCCATTCCGGGTATGG | 100 | 71.8 |
| ebc-r | CTTCCACTGCGGCTGCCAGTT | 100/20/10/5 | 63.4/61.3/60.4/59.5 | |
| fox-uf | blaFOX-1 to blaFOX-6 | Uni-AACATGGGGTATCAGGGAGATG | 100 | 69.6 |
| fox-r | CAAAGCGCGTAACCGGATTGG | 100/20/10/5 | 60.5/58.4/57.5/56.6 | |
| ctx-m-3-uf | blaCTX-M-3 type | Uni-GTTGTTAGGAAGTGTGCCGCTG | 200 | 72.4 |
| ctx-m-3-r | CCTTAGGTTGAGGCTGGGTGAAGT | 200/40/20/10 | 62.3/60.3/59.5/58.7 | |
| ctx-m-9-uf | blaCTX-M-9 type | Uni-TGCAACGGATGATGTTCGCGG | 200 | 73.0 |
| ctx-m-9-r | CCTTTGAGCCACGTCACCAAC | 200/40/20/10 | 60.6/58.5/57.6/56.8 | |
| shv-f | blaSHV type | CCCTCACTCAAGGATGTATTGTGG | 400/80/40/20 | 59.1/57.2/56.4/55.6 |
| shv-ur | Uni-TTAGCGTTGCCAGTGCTCG | 400 | 72.7 | |
| tem-f | blaTEM type | CGCCCTTATTCCCTTTTTTGCGG | 40/8/4/2 | 59.4/57.5/56.8/55.9 |
| tem-ur | Uni-TCAGTGAGGCACCTATCTCAGCG | 40 | 71.3 | |
| Uni | TAMRA-GGTTTCGGATGTTACAGCGT |
a
The primer sequences of six groups of plasmid-mediated AmpC genes were obtained as described previously in reference 28, with minor modifications, and the primer sequences of other genes (blaTEM, blaSHV, blaCTX-M-3, and blaCTX-M-9) were designed in this study. Uni, universal.
b
The concentration-adjusted Tm was calculated according to the nearest-neighbor formula by using the MELTING program (available at http://bioweb.pasteur.fr/seqanal/interfaces/melting.html).