Virulence Constitution of Multi-Drug-Resistant Pseudomonas aeruginosa in Upper Egypt

Figures & data

Table 1 Primers Used in This Study

Target Gene	Sequence (5ʹ – 3ʹ)	Amplicon Size (bp)	Annealing Temperature
toxA	F: 5ʹGGTAACCAGCTCAGCCACAT 3ʹ	352	48°C
	R: 5ʹTGATGTCCAGGTCATGCTTC 3’
plcN	F: 5ʹ GTTATCGCAACCAGCCCTAC 3ʹ	466	48°C
	R: 5ʹAGGTCGAACACCTGGAACAC
plcH	F: 5ʹGAAGCCATGGGCTACTTCAA 3ʹ	307	48°C
	R: 5ʹAGAGTGACGAGGAGCGGTAG 3’
algD	F:5ʹ ATGCGAATCAGCATCTTTGGT 3ʹ	1311	48°C
	R:5ʹ CTACCAGCAGATGCCCTCGGC 3ʹ
lasB	F:5ʹCCAGCCCGCTGACCCACAAGCTGTA 3ʹ	650	46°C
	R: 5ʹCATTCCTTCCTGGAGTGCYRGCCG 3’
aprA	F:5ʹ CCTGATCKGGCCGATAACTGCAAT 3ʹ	1580	46°C
	R:5ʹ GGAAGACASCTATCAATTCGAACAG 3’
exoU	F: 5ʹ GGCACATATCTCCGGTTCCTTC 3ʹ	761	55°C
	R: 5ʹ TCAACTCAGCTGCCAACCATGC 3’
exoS	F: 5ʹATGGCGTGTTCCGAGTCA 3ʹ	1587	55°C
	R: 5ʹ AGGTGTCGGTTCGTGACGTCT 3’
ERIC primers	ERIC-1R, 5VCACTTAGGGGTCCTCGAATGTA-3V
	ERIC-2, 5V-AAGTAAGTGACTGGGGTGAGCG-3V

Abbreviations: toxA, exotoxin A; plcH, hemolytic phospholipase C; plcN, non-hemolytic phospholipase C; algD, GDP-mannose dehydrogenase enzyme for alginate; lasB, elastase; aprA, alkaline protease; exoU, exoenzyme U; exoS, exoenzyme S; ERIC, Enterobacterial Repetitive Intergenic Consensus-PCR.

Table 2 Characteristics of MDR P.aeruginosa Isolates from Different ICUs in Egypt

Isolate No.	toxA	plcN	plcH	algD	lasB	aprA	exoU	exoS	Number of Virulence Genes
PA-M1	-	-	-	-	-	+	+	-	2
PA-M2	-	-	-	-	-	+	+	-	2
PA-M3	-	-	-	-	+	+	-	+	3
PA-M4	-	-	-	-	-	-	-	+	1
PA-M5	-	-	-	-	+	+	+	-	3
PA-M6	-	-	-	-	-	-	+	+	2
PA-M7	-	-	-	-	-	-	+	-	1
PA-M8	-	-	-	-	+	+	-	+	3
PA-M9	-	-	-	-	-	-	-	+	1
PA-M10	-	-	+	+	-	-	+	-	3
PA-M11	-	-	-	+	+	+	+	-	4
PA-M12	-	-	-	-	+	+	-	+	3
PA-M13	-	-	-	-	+	+	+	-	3
PA-M14	-	-	-	-	+	+	+	-	3
PA-M15	+	-	+	+	+	+	-	+	6
PA-M16	-	-	-	-	-	+	+	-	2
PA-M17	+	-	-	+	-	+	-	+	4
PA-M18	+	-	-	+	-	+	+	-	4
PA-M19	+	-	-	+	-	+	-	+	4
PA-M20	-	-	-	-	-	+	+	-	2
PA-M21	-	-	-	-	-	+	+	-	2
PA-M22	-	-	-	-	+	+	-	+	3
PA-M23	-	-	-	-	-	-	-	+	1
PA-M24	-	-	-	-	-	-	-	+	1
PA-M25	-	-	-	-	-	-	+	+	2
PA-M26	-	-	-	-	-	-	+	-	1
PA-M27	+	-	+	-	+	-	+	-	4
PA-M28	-	-	-	-	+	+	+	-	3
PA-M29	-	-	+	+	-	-	+	-	3
PA-M30	-	-	-	+	-	+	+	+	4
PA-M31	-	-	-	-	+	+	-	+	3
PA-M32	+	-	-	+	+	-	-	+	4
PA-M33	-	-	-	-	+	+	+	-	3
PA-M34	+	-	+	+	+	+	-	+	6
PA-M35	-	-	-	-	-	+	+	-	2
PA-M36	-	-	-	-	+	+	+	-	3
PA-M37	-	-	-	-	+	+	-	+	3
PA-M38	+	-	-	+	-	+	-	+	4
PA-M39	+	-	-	+	-	+	-	+	4

Abbreviations: PA, P. aeruginosa; toxA, exotoxin A; plcH, hemolytic phospholipase C; plcN, non-hemolytic phospholipase C; algD, GDP-mannose dehydrogenase enzyme for alginate; lasB, elastase; aprA, alkaline protease; exoU, exoenzyme U; exoS, exoenzyme S; ERIC, Enterobacterial Repetitive Intergenic Consensus-PCR.

Figure 1 Frequency of virulence genes in MDR P. aeruginosa.

Table 3 Virulence Profiles of MDR P. aeruginosa Isolates

Profile	Virulence Gens	Number/%
(Profile 1)	lasB-aprA-exoS	6 (15%)
(Profile 2)	lasB-aprA-exoU	6 (15%)
(Profile 3)	aprA-exoU	6 (15%)
(Profile 4)	toxA-algD-aprA-exoS	5(12.8%)
(Profile 5)	exoS	4(10%)
(Profile 6)	exoU	2 (5%)
(Profile 7)	exoU/exoS	2(5%)
(Profile 8)	toxA-plcH-algD-lasB-aprA-exoS	2(5%)
(Profile 9)	plcH-algD-exoU	2(5%)
(Profile 10)	toxA-algD-aprA-exoU	1(2%)
(Profile 11)	toxA-plcH-lasB-exoU	1(2%)
(Profile 12)	algD-lasB-aprA-exoU	1(2%)
(Profile 13)	algD-aprA-exoU-exoS	1(2%)

Abbreviations: toxA, exotoxin A; plcH, hemolytic phospholipase C; plcN, non-hemolytic phospholipase C; algD, GDP-mannose dehydrogenase enzyme for alginate; lasB, elastase; aprA, alkaline protease; exoU, exoenzyme U; exoS, exoenzyme S; ERIC, Enterobacterial Repetitive Intergenic Consensus-PCR.

Figure 2 Distribution of exoU, exoS genotypes among MDR P. aeruginosa isolates.

Figure 3 Dendrogram illustrating the genetic relationship among MDR P. aeruginosa isolates.