Molecular Identification of Aminoglycoside-Modifying Enzymes and Plasmid-Mediated Quinolone Resistance Genes among Klebsiella pneumoniae Clinical Isolates Recovered from Egyptian Patients

Table 1

Primer sets and PCR cycling conditions used for genotyping and amplification of genes encoding AMES.

Primer	Sequence	Gene	Reference	Amplification conditions	Amplicon size (bp)
aac(3′)-II	F: ATATCGCGATGCATACGCGG; R: GACGGCCTCTAACCGGAAGG	aac(3′)-II	[20]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 55°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	877
aac(6′)-Ib	F: TTGCGATGCTCTATGAGTGGCTA; R: CTCGAATGCCTGGCGTGTTT	aac(6′)-Ib	[20]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 55°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	472
aac(6′)-II	F: CGACCATTTCATGTCC; R: GAAGGCTTGTCGTGTTT	aac(6′)-II	[20]		542
ant(3′′)-I	F: CATCATGAGGGAAGCGGTG; R: GACTACCTTGGTGATCTCG	ant(3′′)-I	[20]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 55°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	787
aph(3′)-VI	F: ATGGAATTGCCCAATATTATT; R: TCAATTCAATTCATCAAGTTT	aph(3′)-VI	[20]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 55°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	780
armA	F: CCGAAATGACAGTTCCTATC; R: GAAAATGAGTGCCTTGGAGG	armA	[20]		846
rmtB	F: ATGAACATCAACGATGCCCTC; R: CCTTCTGATTGGCTTATCCA	rmtB	[20]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 60°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	769
phoE	F: TGGCCCGCGCCCAGGGTTCGAAA; R: GATGTCGTCATCGTTGATGCCGAG	phoE	[21]	Initial denaturation at 95°C for 15 min, then 35 cycles of 95°C for 1 min, 40°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	368
ERIC-1R	R: AACCCACGATGTGGGTAGC	—	[22]		—

acc: aminoglycoside acetyl transferase, ant: aminoglycoside nucleotidyl transferase, aph: aminoglycoside phosphor transferase, arm: aminoglycoside ribosomal methylase, and rmt: ribosomal methylase, phoE; phosphoporin E, ERIC, enterobacterial repetitive intergenic consensus.