Molecular Characteristics and Antibiotic Resistance Profiles of Klebsiella Isolates in Mthatha, Eastern Cape Province, South Africa
Table 1
Primer sequences used for , , , , and genes detection.
Target gene
Primer sequences (5′-3′)
Temp in °C
Target size bp
CTX-M forward primer
ATG TGC AGY ACC AGT AAR GTK ATG GC
58.7
336 bp
CTX-M reverse primer
ATC ACK CGG RTC GCC NGG RAT
59.3
CTX-M1 probe
FAM-CCC gAC AgC Tgg gAg ACg AAA CgT
70.2
CTX-M2 probe (gr 2)
YAK-CAggTgCTTATCgCTCTCgCTCTgTT
66.3
CTX-M9 all/1 probe
LC640-Cg AC AAT ACNgCC ATg AA
41.0
CTX-M9 probe
LC610-CTggATCgCACTgAACCTACgCTgA
53.0
TEM forward primer
AAG TTC TGC TAT GTG CGG TA
59.7
180 bp
TEM reverse primer
TGT TAT CAC TCA TGG TTA TGG CAG C
59.5
SHV forward primer
CAG GAT CTG GTG GAC TAC T
57.2
195 bp
SHV reverse primer
GTC AAG GCG GGT GAC GTT
59.3
SHV-A primer
AAG GCG GGT GAC GTT GTC
59.3
SHV-S primer
CCG GTC AGC GAA AAA CAC
57.0
SHV probe
Cy5-TCT GGC GCA AAA AGG CAG TCA-BBQ
64.5
NDM forward primer
GAC CGC CCA GAT CCT CAA
57.5
51 bp
NDM reverse primer
CGC GAC CGG CAG GTT
60.2
NDM probe
FAM-TGG ATC AAG CA+GGA+GAT-BBQ
48.8
KPC forward primer
GGC CGC CGT GCA ATA C
58,6
61 bp
KPC reverse primer
GCC CAA CTC CTT CA
59,6
KPC probe
Cy5-TGA TAA CGC GCG CAA TTT GT-BBQ
68.9
Relevant positive strains for Klebsiella used as positive control in rPCR: CTX-M (group 1)—E. coli ATCC 25922, CTX-M (all groups except group 1)—CAP2006-D17, TEM—K. pneumoniae ATCC 51503, SHV—K. pneumoniae ATCC 700603, KPC—K. pneumoniae ATCC BAA 1705, NDM—K. pneumoniae ATCC BAA 21246. Obtained from NICD (National Institute of Communicable Diseases, Johannesburg, South Africa).