Characterization of Antibiotic-Susceptibility Patterns, Virulence Factor Profiles and Clonal Relatedness in Proteus mirabilis Isolates from Patients with Urinary Tract Infection in Iran

Figures & data

Table 1 Oligonucleotide Primers Used for PCR Amplification

Gene	Primer Sequence (5′-3′)	Product (bp)	Reference
mrpA	F: ATG AAA TTA AAT AAA TTA GC R: CTG ATA AGT CAG TGC GAA AG	525	This study
mrpH	F: ATG TTT ATA TTT AAA CGA TT R: AGG CAT GGT TAA AAT AAT TG	825	This study
ucaA	F: ATG AAA AGA AAA GTT ATA GC R: CTC ATA GGC AAT GGT GTA AT	540	This study
pmfA	F: CTG CGG CTT TAG TAT TTG GT R: TAA CGG CTT GGA ATT CAC CT	504	This study
ptaA	F: CAA TTT CAG CAC CTA ATA ACC C R: TGC TTA ATC AAG GAG CCG AT	636	This study
zapA	F: TAT CGC AGA ACT GAT CAC TCG R: ATC TGG CTC TTT GTT AGC TTG	540	This study
fliC	F: ATG GCA CAA GTC ATT AAT R: ACG TAA CAG AGA CAG AAC A	1095	This study
ureG	F: AGA ATA TAA TCA ACC ACT GCG TA R: CAT TTT GGC TGT ATC CGC TTC	514	This study
hpmA	F: ATA GTC ACG CCA AAT AAC GAA R: TAT TTC CAC GAG TAG AAC CAG	971	This study
qnrA	F: AGAGGATTTCTCACGCCAGG R: TGCCAGGCACAGATCTTGAC	527	^Citation18
qnrB	F: GGMATHGAAATTCGCCACTG R: TTTGCYGYYCGCCAGTCGAA	264	^Citation18
qnrS	F: GCAAGTTCATTGAACAGGGT R: TCTAAACCGTCGAGTTCGGCG	428	^Citation18
qepA	F: AACTGCTTGAGCCCGTAGAT R: GTCTACGCCATGGACCTCAC	596	^Citation19
aac(6ʹ)-Ib-cr	F: TTGCGATGCTCTATGAGTGGCTA R: CTCGAATGCCTGGCGTGTTT	482	^Citation20

Figure 1 Distribution of P. mirabilis isolates according to the age and gender of UTI patients.

Table 2 Hemagglutination Patterns of the P. mirabilis Isolates

Human Blood Group A		Guinea Pig Erythrocyte		Chicken Erythrocyte
Without mannose	With mannose	Without mannose	With mannose	Without mannose	With mannose
++	++	++	++	+	+

Notes: ++: Strong hemagglutination; +: Weak hemagglutination.

Table 3 Different Patterns of Antibiotic Resistance Among the Isolates

Pattern (P.M)	Antimicrobial Agents	No. of Isolates
P.M1	Sensitive to all	27
P.M2	COT	24
P.M3	AMX, COT	19
P.M4	AMX	9
P.M5	AMX, CAZ, CRO, CPM, ATM, CIP, NOR, TOB, COT	7
P.M6	IMP	3
P.M7	IMP, COT	3
P.M8	AMX, CIP, NOR, TOB, COT	2
P.M9	AMX, IMP, COT	2
P.M10	AMX, CIP	2
P.M11	MEM	2
P.M12	AMX, CAZ, CRO, CPM, ATM, AMC, CIP, NOR, TOB, COT	1
P.M13	AMX, CAZ, CRO, CPM, ATM, TOB, COT	1
P.M14	TOB, COT	1
P.M15	AMX, CAZ, CRO, CPM, ATM, IMP	1
P.M16	AMX, IMP, CIP, NOR, TOB, COT	1
P.M17	AMX, CAZ, MEM, CIP, NOR, COT	1
P.M18	AMX, CAZ, CRO, CPM, ATM, AMC, MEM, IMP, CIP, NOR, TOB, COT	1
P.M19	IMI, CIP, NOR, COT	1
P.M20	AMX, CIP, NOR, COT	1
P.M21	AMX, CAZ, AMC, MEM, IMP	1

Abbreviations: AMX, Amoxicillin; COT, Cotrimoxazole; CAZ, Ceftazidime; CRO, Ceftriaxone; CPM, Cefpodoxime; ATM, Aztreonam; AMC, Amoxicillin-clavulanic acid; CIP, Ciprofloxacin; NOR, Norfloxacin; TOB, Tobramycin; IMP, Imipenem; MEM, Meropenem.

Table 4 Distribution of Antibiotic Resistance According to Inpatients and Outpatients

Antibiotics	Inpatient, n=77 No (%)	Outpatient, n=33 No (%)	Total, n=110 No (%)	P value
AMX	35 (45.5)	14 (42.4)	49 (44.5)	NS
CAZ	13 (16.9)	0	13 (11.8)	NS
CRO	11 (14.3)	0	11 (10)	0.000
CPM	11 (14.3)	0	11 (10)	0.000
ATM	11 (14.3)	0	11 (10)	0.000
AMC	3 (3.9)	0	3 (2.7)	0.000
COT	51 (66.2)	14 (42.2)	65 (59.1)	0.026
TOB	14 (18.2)	0	14 (12.7)	0.000
CIP	17 (22.1)	0	17 (15.5)	0.000
NOR	15 (19.5)	0	15 (13.6)	0.000
IMP	8 (10.4)	5 (15.2)	13 (11.8)	NS
MEM	4 (5.2)	1 (3)	5 (4.5)	NS
MDR	16 (20.8)	0 (0)	16 (14.6)	0.000

Abbreviations: AMX, Amoxicillin; COT, Cotrimoxazole; CAZ, Ceftazidime; CRO, Ceftriaxone; CPM, Cefpodoxime; ATM, Aztreonam; AMC, Amoxicillin-clavulanic acid; CIP, Ciprofloxacin; NOR, Norfloxacin; TOB, Tobramycin; IMP, Imipenem; MEM, Meropenem; NS, Not significant.

Table 5 Relationship Between Antimicrobial Resistance and Biofilm Formation in P. mirabilis Isolates

Antimicrobial Agents
Biofilm Severity	AMX No (%)	CAZ No (%)	CRO No (%)	CPM No (%)	AMC No (%)	MEM No (%)	ATM No (%)	IMP No (%)	CIP No (%)	NOR No (%)	TOB No (%)	COT No (%)
Strong	38 (48.1)	10 (12.7)	9 (11.4)	9 (11.4)	3 (3.8)	3 (3.8)	9 (11.4)	12 (15.2)	14 (17.7)	12 (15.2)	12 (15.2)	45 (57)
Moderate	8 (33.3)	3 (12.5)	2 (8.3)	2 (8.3)	0 (0)	2 (8.3)	2 (8.3)	1 (4.1)	3 (12.5)	3 (12.5)	2 (8.3)	16 (66.7)
Weak	3 (42.9)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	4 (57.1)

Abbreviations: AMX, Amoxicillin; COT, Cotrimoxazole; CAZ, Ceftazidime; CRO, Ceftriaxone; CPM, Cefpodoxime; ATM, Aztreonam; AMC, Amoxicillin-clavulanic acid; CIP, Ciprofloxacin; NOR, Norfloxacin; TOB, Tobramycin; IMP, Imipenem; MEM, Meropenem.

Table 6 Prevalence of Antimicrobial Resistance Among ESBL-Positive and ESBL-Negative Isolates

Antibiotics	ESBL+, n=11 No (%)	ESBL-, n=99 No (%)	Total, n=110 No (%)	P value
AMX	11 (100)	38 (38.4)	49 (44.5)	0.000
CAZ	11 (100)	2 (2)	13 (11.8)	0.000
COT	10 (90.9)	55 (55.6)	65 (59.1)	0.026
TOB	10 (90.9)	4 (4)	14 (12.7)	0.000
CIP	9 (81.8)	8 (8.1)	17 (15.5)	0.000
NOR	9 (81.8)	6 (6.1)	15 (13.6)	0.000
IMP	2 (18.2)	11 (11.1)	13 (11.8)	NS
MEM	1 (9.1)	4 (4)	5 (4.5)	NS
AMC	2 (18.2)	1 (1)	3 (2.7)	0.026
MDR	10 (90.9)	6 (6.1)	16 (14.5)	0.000

Abbreviations: AMX, Amoxicillin; CAZ, Ceftazidime; COT, Cotrimoxazole; TOB, Tobramycin; CIP, Ciprofloxacin; NOR, Norfloxacin; IMP, Imipenem; MEM, Meropenem; AMC, Amoxicillin-clavulanic acid; NS, Not significant.

Figure 2 PFGE results of the selected P. mirabilis isolates. Dendrogram of PFGE was constructed based on UPGMA by using Dice coefficient with a 1.0% band position tolerance. The scores above the dendrogram show the percentage of similarity, and the solid line indicates 80% similarity.

Notes: P1-P4, Pulsotypes 1–4.

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