Characterization of Multidrug Resistant ESBL-Producing Escherichia coli Isolates from Hospitals in Malaysia

Table 1

Primer sequences and PCR conditions.

Primers	Oligonucleotide sequence (5^′ to 3^′)	PCR conditions	Reference	Expected size (bp)
TEM-F TEM-R	ATGAGTATTCAACATTTCCG CTGACAGTTACCAATGCTTA	1 cycle of 5  min at 96^ºC; 35 cycles of 1 min at 96^ºC, 1 min at 58^ºC, 1 min at 72^ºC; 1 cycle of 10 min at 72^ºC	[10]	867
SHV-F SHV-R	GGTTATGCGTTATATTCGCC TTAGCGTTGCCAGTGCTC	1 cycle of 5 min at 96^ºC; 35 cycles of 1 min at 96^ºC, 1 min at 60^ºC, 1 min at 72^ºC; 1 cycle of 10 min at 72^ºC	[10]	867
OXA-F OXA-R	ACACAATACATATCAACTTCGC AGTGTGTTTAGAATGGTGATC	1 cycle of 5 min at 96^ºC; 35 cycles of 1 min at 96^ºC, 1 min at 60^ºC, 2 min at 72^ºC; 1 cycle of 10 min at 72^ºC	[10]	885
CTX-MU1 CTX-MU2	ATGTGCAGYACCAGTAARGT TGGGTRAARTARGTSACCAGA	1 cycle of 7 min at 94^ºC; 35 cycles of 50 sec at 94^ºC, 40 sec at 50^ºC, 1 min at 72^ºC; 1 cycle of 5 min at 72^ºC	[11]	593
DHA-1U DHA-1L	CACACGGAAGGTTAATTCTGA CGGTTATACGGCTGAACCTG	1 cycle of 5 min at 94^ºC; 35 cycles of 30 sec at 94^ºC, 45 sec at 50^ºC, 1 min at 72^ºC; 1 cycle of 8 min at 72^ºC	[12]	970
VEB-1A VEB-1B	CGACTTCCATTTCCCGATGC GGACTCTGCAACAAATACGC	1 cycle of 5 min at 96^ºC; 30 cycles of 1 min at 96^ºC, 1 min at 55^ºC, 2 min at 72^ºC; 1 cycle of 10 min at 72^ºC	[13]	1014
IntI1-F IntI1-R	GGTCAAGGATCTGGATTTGG ACATGCGTGTAAATCATCGTC	1 cycle of 12 min at 94^ºC; 35 cycles of 1 min at 94^ºC, 1 min at 57^ºC, 2 min at 72^ºC; 1 cycle of 10 min at 72^ºC	[14]	500
5^′CS 3^′CS	GGCATCCAAGCAGCAAG AAGCAGACTTGACCTGA	1 cycle of 10 min at 94^ºC; 35 cycles of 1 min at 94^ºC, 1 min at 54^ºC, 2 min at 72^ºC; 1 cycle of 8 min at 72^ºC	[14]	—
IntI2-F IntI2-R	CACGGATATGCGACAAAAAGGT GTAGCAAACGAGTGACGAAATG	Same as for int1	[14]	740
attI2-F orfX-R	GACGGCATGCACGATTTGTA GATGCCATCGCAAGTACGAG	1 cycle of 12 min at 94^ºC; 35 cycles of 1 min at 94^ºC, 1 min at 59.5^ºC, 3.5 min at 72^ºC; 1 cycle of 10 min at 72^ºC	[14]	2000
IntI3-F IntI3-R	AGTGGGTGGCGAATGAGTG TGT TCT TGT ATC GGC AGG TG	1 cycle of 12 min at 94^ºC; 30 cycles of 30 sec at 94^ºC, 30 sec at 60^ºC, 1 min at 72^ºC; 1 cycle of 8 min at 72^ºC	[14]	600
1, 8F 1R 2F 2R 8R 1,8F 9F 9R	GCSATGTGCAGCACCAGTAA ACAAACCGTYGGTGACGATT CTCAATASCGCCATTCCAGG CCGTGGGTTACGATTTTCGC GTCGTACCATAAYCRCCGCT GCSATGTGCAGCACCAGTAA ARTGCAACGGATGATGTYCG GAT GAT TCT CGC CGC TGA AG	1 cycle of 5 min at 95^ºC; 30 cycles of 30 sec at 95^ºC, 30 sec at 63^ºC, and 1 min at 72^ºC; 10 min at 72^ºC.	[15]	666355529846
GES-A GES-B	CTTCATTCACGCACTATTAC TAACTTGACCGACAGAGG	1 cycle of 5 min at 95^ºC; 30 cycles of 1 min at 95^ºC, 45 sec at 55^ºC, 1 min 30 sec at 72^ºC; 1 cycle of 8 min at 72^ºC	[16]	827
VIM-F VIM-R	AGTGGTGAGTATCCGACAG ATGAAAGTGCGTGGAGAC	Same as GES-A	[17]	225
FOXUP1F FOXUP1R	CACCACGAGAATAACC GCCTTGAACTCGACCG	Same as GES-A	[18]	1184
NHAmpCF NHAmpR	ATTCGTATGCTGGATCTCGCCACC CATGACCCAGTTCGCCATATCCTG	Same as GES-A	[18]	396
PER-A PER-B	GGGACARTCSKATGAATGTCA GGGYSGCTTAGATAGTGCTGAT	Same as GES-A
OPAB04	GC ACG CGT T	1 cycle of 2 min 30 sec at 94^ºC; 35 cycles of 30 sec at 94^ºC, 1 min at 47^ºC, and 1 min at 72^ºC; 4 min at 72^ºC.	[7]	—
OPB17	AGGGAACGAG	Same as OPAB04	[8]	—
REP	GCG CCG ICA TGC GGC ATT	1 cycle of 7 min at 94^ºC; 30 cycles of 30 sec at 94^ºC, 1 min at 44^ºC, 8 min at 72^ºC for 30 cycles; 16 min at 65^ºC.	[9]	—
ERIC-1R	ATGTAACGTCCTGGGGATTCAC	1 cycle of 2 min and 30 sec at 94^ºC; 35 cycles of 30 sec at 94^ºC, 1 min at 47^ºC, 1 min at 72^ºC; 1 cycle of 4 min at 72^ºC	[19]	—