TABLE 1

AME primers used in this study

PCRaGeneForward primerReverse primerAmplicon size (bp)
Multiplex 1aac(3)-II5′-CCCTACGAGGAGACTCTGAATG-3′5′-CGAGAATGCCGTTTGAATCGTA-3′442
aac(6′)-II5′-AAGAGTCCGTAACACCGTACAT-3′5′-CCAGAGACTGGTCTATTCCTCG-3′141
Multiplex 2aac(6′)-Ib5′-ACACAATACACAGCATCGTGAC-3′5′-TGTATGGAGTGACGGACTCTTG-3′207
aadA1b5′-ACGATCGACATTGATCTGGCTA-3′5′-CCAAGCGATCTTCTTCTTGTCC-3′343
Multiplex 3aph(3′)-VI5′-TAAAATTGGTCAGTCGCCATCG-3′5′-AAAGCGCTGAAATTGGTTTTGC-3′239
armA5′-GACGAATGAAAGAGTCGCAACA-3′5′-GCTGTTTTAGCACAGGAAGCAT-3′308
rmtB5′-CATAAATCCCCCAAACAGACCG-3′5′-AATCGTACAGGGTATCCAGCTC-3′187
Multiplex 4aadA2b5′-CCGGTTCCTGAACAGGATCTAT-3′5′-CAACTGACTTGATGATCTCGCC-3′319
aadBb5′-CGCAGGTCACATTGATACACAA-3′5′-ATAGTCCAACTCCTCCATGACG-3′212
Uniplexaac(3)-IV5′-GTGCAATACGAATGGCGAAAAG-3′5′-TAGGGAACCTTTGCCATCAACT-3′491
  • a Amplification was done in a 20-μl final volume using the following protocol: denaturation at 95°C for 30 s, followed by 30 cycles consisting of 30 s at 95°C for denaturation, 30 s at 52°C for annealing, and 60 s at 68°C for extension.

  • b Both aadA1 and aadA2 encode ANT(3″)-Ia, and aadB encodes ANT(2″)-Ia (19, 20).