TABLE 2.

Nucleotide sequences of the oligonucleotides used for PCR amplification and DNA sequencing

Gene and primerNucleotide sequence (position)dReference or source
blaTEMa
    JP25′-TTG AAG ACG AAA GGG CCT CGT G-3′ (on promoter 4-25)24
    BLA305′-CTG ACG CTC AGT GGA ACG-3′ (nt 3147-3165 position corresponding to pBR322)c24
    BLA345′-GGG GCC AGA TGG TAA GCC C-3′ (nt 949-968)24
blaSHVb
    SHVA5′-TGG TTA TGC GTT ATA TTC GCC-3′ (nt 120-140)17
    SHVB5′-GGT TAG CGT TGC CAG TGC T-3′ (nt 990-972)17
    SHVC5′-ATC ATG GGA AAG CGT TCA TC-3′ (nt 318-299)This study
    SHVD5′-TTG ATC CGC TCC GTG CTG-3′ (nt 773-790)This study
blaCTX-Mc
    P15′-ATG GTT AAA AAA TCA CTG CGC C-3′ (Y10278; nt 1-22)This study
    P35′-ATG ATG ACT CAG AGC ATT CG-3′ (Y14156; nt 1-20)This study
    P45′-CGG CCT GTA TTT CGC TGT TG-3′ (AF189721; nt 314-333)This study
    P2b5′-TCC CGA CGG CTT TCC GCC TT-3′ (AJ005044; nt 833-814)This study
  • a JP2 and BLA30 (downstream of the 3′ end of the bla gene) were used to amplify the blaTEM genes; JP2, BLA30, and BLA34 were used to sequence the amplified blaTEM genes. The E. coli/pBR322(TEM-1) strain was used as the TEM-specific PCR positive control.

  • b SHVA and SHVB were used to amplify the blaSHV genes; SHVA, SHVB, SHVC, and SHVD were used to sequence the entire range of amplified blaSHV genes. The KPLA-1(SHV-12) strain was used as the SHV-specific PCR positive control.

  • c P1 and P2b were used to amplify blaCTX-M subgroup I genes (including blaCTX-M-1 blaCTX-M-3 and blaCTX-M-10); P3 and P2b were used to amplify blaCTX-M subgroup II genes (including blaCTX-M-2, blaCTX-M-4, blaCTX-M-5, blaCTX-M-6, and blaCTX-M-7); and P4 and P2b were used to amplify blaCTX-M subgroup III genes (including blaCTX-M-8). Sequencing used the same primers as those for PCR. E. coli/pCLL3417(CTX-M-5) was used for the positive control for PCR of subgroup II and the negative control for the other subgroup.

  • d nt, nucleotide.

  • e Downstream of the 3′ end of the bla gene.