Outbreak of Pseudomonas aeruginosa Infections with PER-1 Extended-Spectrum β-Lactamase in Warsaw, Poland: Further Evidence for an International Clonal Complex

DOI: 10.1128/JCM.00997-07

Article Figures & Data

TABLE 1.

Clinical data, PFGE types, MLST results, and β-lactamases corresponding to the P. aeruginosa isolates studied

PFGE type (no. of isolates)	Date(s) (mo/yr) of isolation	Source(s)	Hospital ward(s) or location	MLST^a		β-Lactamase pIs	β-Lactamase variants^b
PFGE type (no. of isolates)	Date(s) (mo/yr) of isolation	Source(s)	Hospital ward(s) or location	Allelic profile	ST	β-Lactamase pIs	β-Lactamase variants^b
A, subtypes 1 to 5 (37)	9/2003-1/2006	Tracheostomy tube swabs, wound swabs, urine, blood, peritoneal fluid, drainage fluid, ear swabs	ICU, surgery, neurology, cardiology, internal medicine	17-5-12-3-14-4-7	ST244	5.3, 7.7, 8.2	PER-1, OXA-2, OXA-50f
B, subtypes 1 and 2 (3)	2/2004-3/2004	Urine, peritoneal fluid, ear swab	Surgery, neurology, nephrology	38-11-3-13-1-2-4	ST235	5.3, 6.5, 7.7, 8.2	PER-1, OXA-2, OXA-74, OXA-50g
C^c (1)	7/2005	Tracheostomy tube swab	Neurology	39-6-12-11-3-15-2	ST245	5.3, 7.7, 8.2	PER-1, OXA-2, OXA-50h
D (2)	2/1992-3/1992		Ankara, Turkey	38-11-3-13-1-2-4	ST235	5.3, 6.5, 7.7, 8.2	PER-1,^c OXA-2,^d OXA-17,^d OXA-50g

↵ a MLST of eight ST244 isolates, three ST235 isolates, one ST245 isolate, and two Turkish isolates was performed.
↵ b PCR analyses of the bla_PER-1-, bla_OXA-2-, bla_OXA-10-, and bla_OXA-50-like genes of all isolates were performed; full sets of amplified genes from three representative ST244 isolates and all ST235 and ST245 isolates from Warsaw were sequenced. Only the bla_OXA-50-like genes from the Turkish isolates were sequenced in the present study; the other β-lactamase genes of these isolates were characterized previously (11, 12).
↵ c Reference 12.
↵ d Reference 11.

TABLE 2.

PCR primers used to analyze β-lactamase genes

Primer designation	Target gene or region	Sequence (5′-3′)	Purpose(s)	Expected size(s) (bp) of amplicon(s) (corresponding primer)^a	Reference
PERA	bla _PER-1	`ATGAATGTCATTATAAAAGC`	PCR, sequencing	925 (PERD)	12
PERD	bla _PER-1	`AATTTGGGCTTAGGGCAGAA`	PCR, sequencing		12
S	bla _OXA-50 type	`AATCCGGCGCTCATCCATC`	PCR, sequencing	867 (AS)	16
AS	bla _OXA-50 type	`GGTCGGCGACTGAGGCGG`	PCR, sequencing		16
OXA-2A	bla _OXA-2	`ATGGCAATCCGAATCTTCG`	PCR, sequencing	828 (OXA-2B)	This work
OXA-2B	bla _OXA-2	`TTATCGCGCAGCGTCCG`	PCR, sequencing		This work
OXA-2C	bla _OXA-2	`CTTGAATGTCGATGCAGGC`	Sequencing		This work
OXA-2D	bla _OXA-2	`GGTCGCAACTGGATACTGC`	Sequencing		This work
OXA-10A	bla _OXA-10 type (bla_OXA-74)	`ATGAAAACATTTGCCGCATATG`	PCR, sequencing	801 (OXA-10B)	This work
OXA-10B	bla _OXA-10 type (bla_OXA-74)	`TTAGCCACCAATGATGCCC`	PCR, sequencing		This work
OXA-10C	bla _OXA-10 type (bla_OXA-74)	`CCTGATGCTCATTCTTTATG`	Sequencing		This work
OXA-10D	bla _OXA-10 type (bla_OXA-74)	`CACCTGAATATCTAGTGCA`	Sequencing		This work
ISPa12.B		`GATCTCGCTTTACATTTACC`	PCR	1,678 (PER-1E)	42
ISPa14.B		`GCCTAATTCGATGCCTTAT`	PCR	1,993 (PER-1E)	42
PER-1E	bla _PER-1	`GCACTGGAACACTAAACTCG`	PCR		This work
PERC	bla _PER-1	`ACACAGCTGTCTGAAACCTC`	PCR	1,883 (ISPa13.A); 2,528 (ISPa14.A)	12
ISPa13.A		`TAACCATATGCACTCAACGG`	PCR		42
ISPa14.A		`AATCAAATGTCCAACCTGCC`	PCR		42

↵ a PCR product sizes are shown in conjunction with the forward primer in each primer pair; the corresponding primer in the pair is identified in parentheses.

TABLE 3.

Nucleotide sequences of identified bla_OXA-50 alleles in the P. aeruginosa isolates studied

↵ a Nucleotide positions are numbered consecutively, starting with the first ATG codon of the bla_OXA-50 coding sequence. Only nucleotide differences with respect to the sequence of the bla_OXA-50 gene from P. aeruginosa PAO1 (GenBank accession no. AE004964 ) (16) are shown. In cases in which polymorphisms cause amino acid differences, the amino acid symbols are given in parentheses.
↵ b GenBank accession no. AY597445 ; reference 25.
↵ c GenBank accession no. AY306131 ; reference 16.

TABLE 4.

Antimicrobial susceptibilities of clinical isolates^a

↵ a Abbreviations: TIC, ticarcillin; TIM, ticarcillin with clavulanate; PIP, piperacillin; TZP, piperacillin-tazobactam; CTX, cefotaxime; CAZ, ceftazidime; CLA, clavulanate; FEP, cefepime; ATM, aztreonam; IPM, imipenem; MEM, meropenem; GEN, gentamicin; TOB, tobramycin; CIP, ciprofloxacin.