Novel Carbapenem-Resistant Klebsiella pneumoniae ST147 Coharboring bla_NDM-1, bla_OXA-48 and Extended-Spectrum β-Lactamases from Pakistan

Figures & data

Table 1 Primer Sequences Used for PCR

Gene	Primer Sequence (5ʹ–3ʹ)	Annealing (°C)	Amplicon Size (bp)	Reference
rpoB (K. pneumoniae)	F: CAA CGG TGT GGT TAC TGA CG R: TCT ACG AAG TGG CCG TTT TC	55	108	^Citation53
pehX (K. oxytoca)	F: GAT ACG GAG TAT GCC TTT ACG GTG R: TAG CCT TTA TCA AGC GGA TAC TGG	55	343	^Citation53
gyrA (Klebsiella genus)	F: CGC GTA CTA TAC GCC ATG AAC GTA R: ACC GTT GAT CAC TTC GGT CAG G	55	441	^Citation53
blaSHV	F: CTT TAT CGG CCC TCA CTC AA R: AGG TGC TCA TCA TGG GAA AG	55	237	^Citation54
blaTEM	F: CGC CGC ATA CAC TAT TCT CAG AAT GA R: ACG CTC ACC GGC TCC AGA TTT AT	55	445	^Citation54
blaCTX-M	F: ATG TGC AGY ACC AGT AAR GTK ATG GC R: TGG GTR AAR TAR GTS ACC AGA AYC AGC GG	55	593	^Citation54
blaNDM-1	F: ATG GAA TTG CCC AAT ATT ATG CAC R: TCA GCG CAG CTT GTC GGC	52	813	^Citation55
blaVIM	F: GAT GGT GTT TGG TCG CAT A R: CGA ATG CGC AGC ACC AG	52	390	^Citation56
blaOXA-48	F: GCG TGG TTA AGG ATG AAC AC R: CAT CAA GTT CAA CCC AAC CG	52	438	^Citation56
blaIMP	F: GGA ATA GAG TGG CTT AAY TCT C R: GGT TTA AYA AAA CAA CCA CC	52	232	^Citation56
blaNDM-1	F: TGGCTTTTGAAACTGTCGCACC R: CTGTCACATCGAAATCGCGCGA	60	1000	^Citation57
gapA	F: TGA AGT ATG ACT CCA CTC ACG G R: AAC GCC TTT CAT TGC GCC TTC GGA A	60	662	^Citation20
infB	F: CTC TCT GCT GGA CTA CAT TCG R: CGC TTT CAG CTC CAG AAC TTC	52	462	^Citation20
mdh	F: CCC AAC TGC CTT CAG GTT CAG R: CCT TCC ACG TAG GCG CAT TCC	52	756	^Citation20
pgi	F: GAG AAA AAC CTG CCG GTG CTG CTG R: CGG TTA ATC AGG CCG TTA GTG GAG C	52	566	^Citation20
phoE	F: ACC TGG CGC AAC ACC GAT TTC TTC R: TTC AGC TGG TTG ATT TTG TAA TCC AC	52	602	^Citation20
rpoB	F: GGC GAA ATG GCG GAA AAC CA R: GAG TCT TCG AAG TTG TAA CC	52	1075	^Citation20
tonB	F: CTC TAT ACT TCG GTA CAT CAG GTT R: CCT GTT TGG CGG CCA GCA CCT GGT	48	539	^Citation20

Abbreviations: rpoB, RNA polymerase beta-subunit gene; pehX, polygalacturonase gene; gyrA, DNA gyrase subunit A gene; blaSHV, beta-lactamase sulfhydryl reagent variable gene; blaNDM, New Delhi metallo-beta-lactamase gene; blaVIM, metallo-beta-lactamase verona integron gene; blaTEM, beta-lacatmase temoneira gene; blaOXA-48, beta-lactamase oxacillinase 48 gene; blaCTX-M, beta-lactamase cefotaxime munich gene; blaIMP, beta-lactamase imipenemase gene; gapA, glyceraldehyde-3-phosphate dehydrogenase A gene; infB, translation initiation factor IF-2 gene; mdh, malate dehydrogenase gene; pgi, phosphoglucose isomerase gene; phoE, phosphoporin E gene; tonB, periplasmic energy transducer gene; rpoB, beta-subunit of RNA polymerase gene.

Table 2 MIC Values of Selected Carbapenem-Resistant K. pneumoniae Strains

Strain ID	Resistance Profile	MIC (μg/mL)
		MEM	CAZ	SXT	AMP	AMC	CPT	CIP	ATM	AK	PB	DO	CTX
KP-17	MDR	4	8	2/38	64	32/16	0.5	1	16	16	2	4	1
KP-97	XDR	512	32	8/152	8	8/4	16	8	32	8	8	16	8
KP-104	MDR	64	32	2/38	8	16/8	0.5	1	4	64	2	4	8
KP-188	MDR	32	64	2/38	16	32/16	0.5	1	4	16	2	4	32
KP-191	MDR	16	4	16/304	8	32/16	0.5	1	8	16	2	2	16
KP-194	MDR	16	4	4/76	16	8/4	0.5	1	4	32	2	2	8
KP-199	XDR	>1024	128	32/608	512	256/128	0.5	64	128	128	8	32	256
KP-222	MDR	256	128	2/38	64	32/16	0.5	1	4	8	2	4	32
KP-246	XDR	128	256	8/152	32	64/32	16	1	16	64	16	64	64
KP-268	XDR	512	256	16/304	128	16/8	16	64	64	1024	4	128	256
KP-272	MDR	256	64	2/38	8	8/4	0.5	1	32	64	2	4	32
KP-284	MDR	64	32	2/38	32	8/4	0.5	16	4	16	2	4	32
KP-289	XDR	64	16	8/152	32	256/128	32	128	128	512	8	512	8
KP-315	XDR	>1024	256	8/152	64	128/64	16	4	512	512	8	256	4
KP-326	MDR	128	64	2/38	16	8/4	0.5	0.5	32	16	2	4	16
KP-333	MDR	32	16	8/152	8	8/4	0.5	0.5	16	16	2	4	2
KP-426	MDR	8	4	2/38	16	16/8	0.5	1	16	32	2	4	0.5
KP-443	MDR	64	32	4/76	8	128/64	0.5	1	16	8	2	4	0.5
KP-465	MDR	128	128	2/38	16	8/4	0.5	1	128	16	32	4	0.5
KP-494	MDR	64	128	2/38	8	8/4	4	32	64	16	2	4	0.5
KP-544	XDR	32	64	32/608	32	128/64	0.5	1	256	512	16	128	256
KP-562	MDR	64	256	2/38	64	8/4	0.5	1	32	128	2	4	0.5
KP-611	XDR	256	64	64/1216	64	128/64	0.5	16	16	64	2	4	64
KP-663	MDR	128	512	32/608	64	8/4	0.5	1	64	8	2	4	0.5
KP-675	MDR	32	32	2/38	256	8/4	0.5	1	64	8	8	4	0.5
KP-668	MDR	64	32	2/38	8	64/32	2	1	128	16	1	4	0.5
KP-687	MDR	32	64	2/38	128	8/4	8	1	32	16	1	4	0.5
KP-704	XDR	512	128	64/1216	128	256/128	0.5	256	32	1024	16	512	32

Abbreviations: MDR, multidrug resistant; XDR, extensively drug resistant.

Table 3 Resistance Profile of Carbapenem-Resistant K. pneumoniae

Strain ID	Ward	Sample Type	MIC (μg/mL) Imipenem	Resistance Profile	Profile of Resistance Genes	Replicon and Sequence Type
KP-17	SICU	Wound	256	MDR	blaNDM-1 (MT312213)*, blaOXA-48, blaCTX-M, blaSHV	ST147, IncL/M, IncFII, IncA/C, IncFH1
KP-97	SICU	Pus	64	XDR	blaNDM-1 (MT320894)*, blaCTX-M, blaSHV	-
KP-104	OPD	Wound	32	MDR	blaNDM-1 (MT320895)*, blaCTX-M, blaSHV	-
KP-188	SICU	Tracheal secretion	64	MDR	blaNDM-1 (MT320896)*, blaSHV	-
KP-191	GS	Wound	16	MDR	blaNDM-1 (MT320897)*, blaCTX-M	-
KP-194	GS	Wound		MDR	blaNDM-1 (MT320898)*, blaSHV	-
KP-199	OPD	Pus	>1024	XDR	blaNDM-1 (MT320899)*, blaOXA-48, blaCTX-M, blaSHV	ST147, IncL/M, IncFII, IncA/C
KP-222	GS	Sputum	128	MDR	blaNDM-1 (MT320900)*, blaOXA-48, blaCTX-M	ST147, IncL/M, IncFII, IncA/C, IncFH1
KP-246	GS	Wound	32	XDR	blaNDM-1 (MT320901)*, blaCTX-M	-
KP-268	SICU	Tip cells	64	XDR	blaNDM-1 (MT320902)*, blaTEM, blaCTX-M, blaSHV	-
KP-272	GS	Wound	32	MDR	blaNDM-1 (MT320903)*, blaCTX-M, blaSHV	-
KP-284	GS	Wound	8	MDR	blaNDM-1 (MT320904)*, blaCTX-M, blaSHV	-
KP-289	GS	Wound	16	XDR	blaNDM-1 (MT320905)*, blaTEM	-
KP-315	SICU	Tracheal secretion	32	XDR	blaNDM-1 (MT320906)*, blaSHV	-
KP-326	OPD	Blood	128	MDR	blaNDM-1 (MT320907)*, blaOXA-48, blaCTX-M	ST147, IncL/M, IncFII, IncA/C, IncFH1
KP-333	GS	Pus	32	MDR	blaNDM-1 (MT320908)*, blaSHV	-
KP-426	BURN	Tracheal secretion	64	MDR	blaNDM-1 (MT320909)*, blaCTX-M	-
KP-443	GS	Wound	64	MDR	blaNDM-1 (MT320910)*	-
KP-465	WSW	Wound	128	MDR	blaNDM-1 (MT320911)*, blaOXA-48, blaSHV	ST11, IncL/M, IncFII, IncA/C, IncH1
KP-494	SICU	Tracheal secretion	64	MDR	blaNDM-1 (MT320912)*, blaOXA-48, blaCTX-M, blaSHV	ST147, IncL/M, IncFII, IncA/C
KP-544	GS	Blood	32	XDR	blaNDM-1 (MT320913)*	-
KP-562	SICU	Tracheal secretion	64	MDR	blaNDM-1 (MT320914)*, blaCTX-M	-
KP-611	PS	Pus	128	XDR	blaNDM-1 (MT320915)*, blaCTX-M	-
KP-663	GS	Wound	64	MDR	blaNDM-1 (MT320916)*, blaSHV, blaCTX-M	-
KP-675	SICU	Tracheal secretion	128	MDR	blaNDM-1 (MT320917)*, blaOXA-48, blaCTX-M	ST11, IncL/M, IncFII, IncA/C
KP-668	GS	Pus	64	MDR	blaNDM-1 (MT320918)*, blaTEM, blaSHV	-
KP-687	SICU	Tracheal secretion	32	MDR	blaNDM-1 (MT320919)*, blaSHV	-
KP-704	CM	Sputum	32	XDR	blaNDM-1 (MT320920)*, blaCTX-M	-

Note: *GenBank Accession Number.

Abbreviations: GS, general surgery; PS, plastic surgery; SICU, surgical ICU; CM, chest medicine.

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