Prevalence, Antimicrobial Resistance, and Molecular Description of Pseudomonas aeruginosa Isolated from Meat and Meat Products

Table 1

PCR conditions [26–30].

Molecular characters	Targeted genes	Primer sequence (5′-3′)	PCR product (bp)	PCR programs	PCR volume (50 μL)
Virulence factors	algD	F: AAG GCG GAA ATG CCA TCT CC	275	1 cycle: 2 min at 95°C; 30 cycles: 30 s at 94°C; 30 s at 58°C; 60 s at 72°C; 1 cycle: 7 min at 72°C	dNTP: 200 μM; PCR buffer (10X): 5 μL; Primer F: 0.5 μM; Primer R: 0.5 μM; Mgcl2: 1.5 mM; DNA: 2.5 μL; Taq DNA polymerase: 1.25 U
		R: AGG GAA GTT CCG GGC GTT TG
	algU	F: CGC GAA CCG CAC CAT CGC TC	410
		R: GCC GCA CGT CAC GAG C
	exoT	F: CAA TCA TCT CAG CAG AAC CC	1159	1 cycle: 5 min at 96°C; 30 cycles: 30 s at 94°C; 30 s at 47–63 ^0C; 1 min at 72°C; 1 cycle: 3 min at 72°C
		R: TGT CGT AGA GGA TCT CCT G
	lasA	F: GCA GCA CAA AAG ATC CC	1075
		R: GAA ATG CAG GTG CGG TC
	pvdA	F: GAC TCA GGC AAC TGC AAC	1281
		R: TTC AGG TGC TGG TAC AGG
	lasB	F: ACA CAA TAC ATA TCA ACT TCG C	284	1 cycle: 3 min at 94°C; 30 cycles: 30 s at 94°C; 60 s at 55°C; 90 s at 72°C; 1 cycle: 5 min at 72°C
		R: AGT GTG TTT AGA ATG GTG ATC
	toxA	F: ATG TGC AGY ACC AGT AAR GT	270
		R: TGG GTR AAR TAR GTS ACC AGA
	plcH	F: CAC ACG GAA GGT TAA TTC TGA	608
		R: CGG TTA RAC GGC TGA ACC TG
	plcN	F: CGA CTT CCA TTT CCC GAT GC	481
		R: GGA CTC TGC AAC AAA TAC GC
	exoS	F: GTG TGC TTT ATG CCA TGA G	444
		R: GGT TTC CTT TTC CAG GTC
	exoY	F: TAT CGA CGG TCA TCG TCA GGT	1035	1 cycle: 5 min at 96°C; 30 cycles: 30 s at 94°C; 30 s at 47–63 ^0C; 1 min at 72°C; 1 cycle: 3 min at 72°C
		R: TTG ATG CAC TCG ACC AGC AAG
	exoU	F: GAT TCC ATC ACA GGC TCG	3308
		R: CTA GCA ATG GCA CTA ATC G
	apr	F: TGT CCA GCA ATT CTC TTG C; R: CGT TTT CCA CGG TGA CC	1017
	phzM	F: ATG GAG AGC GGG ATC GAC AG	875
		R: ATG CGG GTT TCC ATC GGC AG
	phzS	F: TCG CCA TGA CCG ATA CGC TC	1752
		R: ACA ACC TGA GCC AGC CTT CC
	phzH	F: GCC AAG GTT TGT TGT CGG	1036
		R: CGC ATT GAC GAT ATG GAA C
	pilA	F: ACA GCA TCC AAC TGA GCG	1675
		R: TTG ACT TCC TCC AGG CTG
	pilB	F: TCG AAC TGA TGA TCG TGG	408
		R: CTT TCG GAG TGA ACA TCG
Antibiotic resistance genes	blaTEM^a	F: ATG AGT ATT CAA CAT TTC CG	747	1 cycle: 5 min at 96°C; 35 cycles: 60 s at 96°C; 60 s at 58°C; 60 s at 72°C; 1 cycle: 10 min at 72°C
		R: CTG ACA GTT ACC AAT GCT TA
	blaSHV^b	F: GGT TAT GCG TTA TAT TCG CC	867	1 cycle: 5 min at 96°C; 35 cycles: 60 s at 96°C; 60 s at 60°C; 60 s at 72°C; 1 cycle: 10 min at 72°C
		R: TTA GCG TTG CCA GTG CTC
	blaOXA^c	F: ACA CAA TAC ATA TCA ACT TCG C	885
		R: AGT GTG TTT AGA ATG GTG ATC
	blaCTX-M^d	F: ATG TGC AGY ACC AGT AAR GT	593	1 cycle: 7 min at 94°C; 35 cycles: 50 s at 94°C; 40 s at 50 ^0C; 60 s: At 72°C; 1 cycle: 5 min at 72°C
		R: TGG GTR AAR TAR GTS ACC AGA
	blaDHA^e	F: CAC ACG GAA GGT TAA TTC TGA	970	1 cycle: 5 min at 94°C; 35 cycles: 30 s at 94°C; 45 s at 50°C; 60 s at 72°C; 1 cycle: 8 min at 72°C
		R: CGG TTA RAC GGC TGA ACC TG
	blaVEB^f	F: CGA CTT CCA TTT CCC GAT GC	1014	1 cycle: 5 min at 96°C; 30 cycles: 60 s at 96°C; 60 s at 55°C; 2 min at 72°C; 1 cycle: 10 min at 72°C
		R: GGA CTC TGC AAC AAA TAC GC
	gyrA	F: GTG TGC TTT ATG CCA TGA G	287	1 cycle: 14 min at 95°C; 35 cycles: 45 s at 95°C; 45 s at 51°C; 60 s at 71°C; 1 cycle: 7 min at 71°C
		R: GGT TTC CTT TTC CAG GTC
	parC	F: CAT CGT CTA CGC CAT GAG	267
		R: AGC AGC ACC TCG GAA TAG

Annealing temperatures of exoT, exoY, exoU, apr, phzM, phzS, phzH, lasA, lasB, pvdA, pilA, and pilB virulence factors were 54, 61.8, 61.6, 51, 54, 63, 51, 47, 57, 59, 59, and 54°C, respectively. ^aTemoneira: the name of the first patient providing the first sample. ^bSulfhydryl reagent variable. ^cActive on oxacillin. ^dActive on cefotaxime, first isolated at Munich. ^eDiscovered at Dhahran, Saudi Arabia. ^fVietnam extended-spectrum β-lactamase.