Table 4.
Primers used for screening for β-lactamase genes
| Target Gene | Primer name | 5'-3' sequence | T°C | Size (bp) | Gene accession number |
|---|---|---|---|---|---|
| blaTEM |
TEM-F |
ATGAGTATTCAACAT TTC CG |
55 |
840 |
EF125012-related |
| TEM-R |
CCAATGCTTAATCAG TGA GG |
||||
| blaSHV |
SHV-F |
TTCGCCTGTGTATTATCTCCCTG |
50 |
854 |
AF148850-related |
| SHV-R |
TTAGCGTTGCCAGTGYTCG |
||||
| blaCTX-M concensus |
MA1 |
ATGTGCAGYACCAGTAARGTKATGGC |
60 |
593 |
Y10278-related |
| MA2 |
TGGGTRAARTARGTSACCAGAAYCAGCGG |
||||
| CTX-M group I |
CTXM1-F3 |
GAC GAT GTC ACT GGC TGA GC |
55 |
499 |
X92506-related |
| CTXM1-R2 |
AGC CG C CGA CGC TAA TAC A |
||||
| CTX-M group II |
TOHO1-2 F |
GCG ACC TGG TTA ACT ACA ATC C |
55 |
351 |
X92507-related |
| TOHO1-1R |
CGG TAG TAT TGC CCT TAA GCC |
||||
| CTX-M group III |
CTXM825F |
CGC TTT GCC ATG TGC AGC ACC |
55 |
307 |
AF189721-related |
| CTXM825R |
GCT CAG TAC GAT CGA GCC |
||||
| CTX-M group IV |
CTXM914F |
GCT GGA GAA AAG CAG CGG AG |
62 |
474 |
AF252622-related |
| CTXM914R |
GTA AGC TGA CGC AAC GTC TG |
||||
| blaCMY (consensus) |
CF1 |
ATGATGAAAAAATCGTTATGC |
55 |
1200 |
U77414-related |
| CF2 |
TTGCAGCTTTTCAAGAATGCGC |
||||
| blaCMY-1 group |
CMY-1 F |
GTGGTGGATGCCAGCATCC |
58 |
915 |
AJ291609-related |
| CMY-1R |
GGTCGAGCCGGTCTTGTTGAA |
||||
| blaCMY-2 group |
CMY-2 F |
GCACTTAGCCACCTATACGGCAG |
58 |
758 |
AF305559-related |
| CMY-2R |
GCTTTTCAAGAATGCGCCAGG |
||||
| blaOXA-1 |
OXA-1 F |
ATGAAAAACACAATACATATCAACTTCGC |
62 |
820 |
JO2967-related |
| OXA-1R |
GTGTGTTTAGAATGGTGATCGCATT |
||||
| blaOXA-2 |
OXA-2 F |
ACGATAGTTGTGGCAGACGAAC |
62 |
602 |
AF300985-related |
| OXA-2R |
ATYCTGTTTGGCGTATCRATATTC |
||||
| blaPER-concensus |
PER-F |
ATGAATGTCATTATAAAAGC |
55 |
925 |
Z21957-related |
| PER-R |
AATTTGGGCTTAGGGCAGAA |
||||
| blaACC-like |
ACC-F |
AGCCTCAGCAGCCGGTTAC |
53 |
818 |
AJ133121-related |
| ACC-R |
GAAGCCGTTAGTTGATCCGG |
||||
| blaVEB-concensus |
VEB-F |
ATTTAACCAGATAGGACTACA |
55 |
1000 |
Z21957-related |
| VEB-R |
CGGTTTGGGCTATGGGCAG |
||||
| blaDHA-concensus |
DHA-F |
TGATGGCACAGCAGGATATTC |
55 |
997 |
EF406115-related |
| DHA-R | GCTTTGACTCTTTCGGTATTCG |
Primer combinations used for screening and sequencing bla genes. The consensus primers were used for screening and sequencing purposes except for blaCTX-M, and blaOXA that were sequenced using group-specific primers. CTX-M group I primers detect genes encoding CTX-M −1, -3, -10 to −12, -15, -22,- 23, -28, -29 and −30 while primers for CTX-M group II primers detect genes encoding CTX-M-2, -4, -7, and −20. Group III primers detect only CTX-M-8 while group IV primers detect genes encoding CTX-M-9, -13, -14, -16 to 19, -21, and 27.
T°C: annealing temperature.
Y = T or C, R = G or A, S = G or C, K = G or T.