Detection of pap, sfa and afa adhesin-encoding operons in uropathogenic Escherichia coli strains: Relationship with expression of adhesins and production of toxins

Blanco, Miguel; Alonso, M. P.; Mora, Azucena; Balsalobre, Carlos; Muñoa, F.; Juárez, Antonio; Blanco, Jesús E.

doi:10.1016/s0923-2508(97)82450-3

Cited by 83 publications

(67 citation statements)

References 34 publications

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“…Bacterial DNA to be amplified was released from whole organisms by boiling, and a multiplex PCR assay was carried out in a total volume of 25 µL, containing 2 µL of template DNA, each of the primers at 60 ng, the four deoxynucleotide triphosphates (each at 200 µM), PCR buffer 1X and 1.5 U of Taq DNA polymerase 3,10 . For Multiplex PCR, amplifications consisted of 25 cycles of 94 ºC for one min, specific annealing temperature for each primer for one min (Table1), 68 ºC for three min, and a final extension at 72 ºC for seven min in a Thermal Cycler (Gene Amp PCR System 9700/Perkin Elmer Corporation, Norwal CT/USA).…”

Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Genotypic characterization of virulence factors in Escherichia coli strains from patients with cystitis

Tiba

Yano

Leite

2008

Rev. Inst. Med. trop. S. Paulo

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Abbreviations: aerobactin (iucD), afimbrial adhesin (afaB/C), cytolethal distending toxin (cdtB), cytotoxic necrotizing factor type 1 (cnf-1), G adhesin classes of P fimbriae (papG alleles), group II capsule (kpsMTII), α-hemolysin (hly), minor structural subunits of P fimbriae (papE/F), outer membrane protein of P fimbrae (papC), polymerase chain reaction (PCR), S fimbriae (sfaC/D), type 1 fimbriae (fimH), urinary tract infection (UTI), uropathogenic specific protein (usp), uropathogenic Escherichia coli (UPEC), virulence factors (VFs SUMMARYAdhesins (P-fimbriae, S-fimbriae, type 1 fimbriae and afimbrial adhesin), toxins (α-hemolysin and cytotoxic necrotizing factor type 1), iron acquisition systems (aerobactin) and host defense avoidance mechanisms (capsule or lipopolysaccharide) have been shown to be prevalent in Escherichia coli strains associated with urinary tract infections. In this work, 162 Uropathogenic Escherichia coli (UPEC) strains from patients with cystitis were genotypically characterized by polymerase chain reaction (PCR) assay. We developed three multiplex PCR assays for virulence-related genes papC, papE /F, papG alleles, fimH, sfa/foc, afaE, hly, usp, cdtB, iucD, and kpsMTII, all

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Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Genotypic characterization of virulence factors in Escherichia coli strains from patients with cystitis

Tiba

Yano

Leite

2008

Rev. Inst. Med. trop. S. Paulo

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“…E. coli adhesion to host cells is important to bacteria infection and persistence in urinary fluxes (6,10). The genes involved in biosynthesis of fimbria and adhesins present in UPEC are organized in operons denominated pap and sfa, coding for P and S fimbrial adhesins (2,3,24). IRGA homologue adhesin (IHA) is an outer membrane protein (OMP) found mainly in UPEC being also involved in adherence (13).…”

mentioning

confidence: 99%

“…The primers and PCR conditions were previously described (1,2,4,7,12,15,16). Amplicons identities were confirmed by sequencing (Amersham Pharmacia Biotech).…”

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confidence: 99%

Virulence factors and antimicrobial resistance of escherichia coli isolated from urinary tract of swine in southern of Brazil

Costa¹,

Drescher²,

Maboni³

et al. 2008

Braz. J. Microbiol.

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The present study determined the molecular and resistance patterns of E. coli isolates from urinary tract of swine in Southern of Brazil. Molecular characterization of urinary vesicle samples was performed by PCR detection of virulence factors from ETEC, STEC and UPEC. From a total of 82 E. coli isolates, 34 (38.63%) harbored one or more virulence factors. The frequency of virulence factors genes detected by PCR were: pap (10.97%), hlyA (10.97%), iha (9.75%), lt (8.53%), sta (7.31%) sfa (6.09%), f4 (4.87%), f5 (4.87%), stb (4.87%), f6 (1.21%) and f41 (1.21%). Isolates were resistant to penicillin (95.12%), lincomycin (93.9%), erythromycin (92.68%), tetracycline (90.24%), amoxicillin (82.92%), ampicillin (74.39%), josamycin (79.26%), norfloxacin (58.53%), enrofloxacin (57.31%), gentamicin (39.02%), neomycin (37.8%), apramycin (30.48%), colistine (30.48%) and cefalexin (6.09%). A number of 32 (39.02%) E. coli isolates harbored plasmids.

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“…The E. coli samples were genotyped using two mPCR assays to detect ϐimbria and toxins by amplifying the following gene encoding for regions: STa (158 bp), STb (113 bp), LT (272 bp), STx (758 bp), F4 (499 bp), F5 (230 bp), F6 (520 bp), F18 (313 bp) and F41 (613 bp), for the intestinal, fecal and tissue samples; and cnf (543 bp), hly (1117 bp), bfp (326 bp), eae (368 bp), pap (328 bp), iha (827 bp), sfa (410 bp), cnf (543 bp) and usp (440 bp) for the urinary isolates, as previously described (Brito et al 2004, Costa et al 2008, Blanco et al 1996, Blanco et al 1997 Aliquots of 10μl ampliϐication products were subjected to electrophoresis for 45 minutes at 100V in 1.5% agarose gel stained with ethidium bromide. DNA fragments were compared with a 50 bp DNA ladder.…”

Section: Methodsmentioning

confidence: 99%

Phylogenetic and pathotype analysis of Escherichia coli swine isolates from Southern Brazil

et al. 2012

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The current study evaluated the presence of virulence factors by a multiplex PCR technique and then phylogenetically classified the studied strains into groups A, B1, B2 and D, according to Clermont et al. (2000), in 152 intestinal and extraintestinal swine isolates of Escherichia coli. Seventy seven isolates tested were positive for virulence factors. Phylogenetic characterization placed 21 samples into group A, 65 into B1, 19 into B2 and 47 into D. Fourteen urine samples were classified as uropathogenic E. coli (UPEC), nine were both UPEC and enterotoxigenic E. coli (ETEC) and four were ETEC only. The most common phylogenetic classifications were B1 and D groups. Of the analyzed fecal samples, 25 were classified as ETEC. Phylogenetically, the group of higher occurrence was B1, followed by B2, A and D. For the small intestine samples, 20 were classified as ETEC. Phylogenetic analysis found groups B1 and A to be the most commons in these samples. Six isolated tissue samples were classified as ETEC and most of them were designated as group D by phylogenetic classification. The phylogenetic analysis could be employed in veterinary laboratories in the E. coli isolates screening, including the possibility of vaccine strain selection and epidemiological searches.

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Detection of pap, sfa and afa adhesin-encoding operons in uropathogenic Escherichia coli strains: Relationship with expression of adhesins and production of toxins

Cited by 83 publications

References 34 publications

Genotypic characterization of virulence factors in Escherichia coli strains from patients with cystitis

Genotypic characterization of virulence factors in Escherichia coli strains from patients with cystitis

Virulence factors and antimicrobial resistance of escherichia coli isolated from urinary tract of swine in southern of Brazil

Phylogenetic and pathotype analysis of Escherichia coli swine isolates from Southern Brazil

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