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  • 1
    Electronic Resource
    Electronic Resource
    Cloning of a Vero toxin (VT2) gene from a VT2-converting phage isolated from Escherichia coli 0157: H7 (1987)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 48 (1987), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract A Vero toxin (VT2 or Shiga-like toxin II)-converting phage was isolated from Escherichia coli 0157: H7 strain J-2. Nontoxigenic E. coli C600 produced VT2 when lysogenized with the toxin-converting phage. EcoRI fragments of the phage DNA were ligated with EcoRI-digested pBR322 or pUC118 and were transformed into E. coli MC1061 or MV1184. Transformants exhibiting VT2 production commonly contained a 4.6 kb EcoRI fragment. It was found that a 2.3 kb KpnI-SphI fragment coded VT2 production and that this fragment hybridized weakly with the 2.1 kb fragment encoding VT1.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1987.tb02555.x
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  • 2
    Electronic Resource
    Electronic Resource
    Procedures for large-scale production and purification of Clostridium botulinum C1 toxin for preparation of toxoid (1985)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 30 (1985), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Fortified cooked meat medium containing calcium carbonate (CaCO3-FCM) supported toxin production of a strain of Clostridium botulinum type C to a level of 2 × 106 mouse i.p. LD50/ml. C1 toxin was purified by sequential steps of acid precipitation from 5-fold diluted culture supernatant in the presence of RNA, 2nd acid precipitation by dialysis, removal of RNA by protamine treatment, removal of excess protamine and bufferisation by ultrafiltration through Amicon PM-30 membrane, sulphopropyl-Sephadex chromatography, and Sephadex G-200 gel filtration. By these procedures, 25 mg or more of highly purified C1 toxin was constantly obtained from a lot of 600-ml culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1985.tb00983.x
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  • 3
    Electronic Resource
    Electronic Resource
    Structural and sequence diversity of the pathogenicity island of uropathogenic Escherichia coli which encodes the USP protein (2001)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 205 (2001), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A total of 321 uropathogenic Escherichia coli (UPEC) strains and 12 strains of E. coli isolated from stool samples of healthy individuals, which were previously shown to be positive in colony hybridization test using the usp (encoding for the uropathogenic-specific protein) DNA probe, were examined by PCR amplification to determine the size of the usp gene and the pathogenicity island (PI). Three types of size variation were observed for the usp gene and four types for the PI. Sequencing analysis of the PIs from seven representative strains (six UPEC and one from a normal healthy individual) revealed that the usp genes can be classified into two groups, each having different sequences in the 3′-terminal region. The peptides encoded by the three open reading frames (ORFs) downstream of usp had identical 23 amino acid residues in the C-terminal region. The subregion encoding these small ORFs has a mosaic structure constituted of six segments. The positions of these segments vary from strain to strain, and in some strains, two to four segments are deleted. This indicates that rearrangements occur frequently in this region and the mosaic arrangement apparently contributes to the size variation observed in the PCR examination of the usp genes and PIs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.2001.tb10927.x
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  • 4
    Electronic Resource
    Electronic Resource
    Detection of Proteus mirabilis Urease Gene in Urinary Calculi by Polymerase Chain Reaction (1996)
    Oxford, UK : Blackwell Publishing Ltd
    International journal of urology 3 (1996), S. 0 
    Details
    ISSN: 1442-2042
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background: Urea-splitting microorganisms cannot always be detected by stone or urine culture in patients with infection stones. Detection of genetic elements within the calculi by the polymerase chain reaction (PCR) may be a useful alternative. In this study, we assessed the usefulness of the PCR method in detecting the urease gene specific to Proteus mirabilis in urinary calculi.Methods: Thirty-eight metabolic stones (calcium oxalate and/or calcium phosphate, uric acid, or cystine) and 49 struvite stones were examined. The PCR was applied with DNA extracted by boiling pulverized stone pieces.Results: Of the 87 stones, PCR demonstrated the presence of the P. mirabilis urease elements ureC1 and ureC2 in 1 7, all of which were struvite. Stone culture and urine culture had been performed in 22 and 46 struvite stone cases, respectively, and the PCR was positive in all of the 10 culture-positive calculi and also in two calculi from which P. mirabilis was not isolated.Conclusion: PCR was reliable and convenient for detecting P. mirabilis in desiccated struvite calculi. Study to detect other species such as Ureaplasma or Corynebacterium would be useful in elucidating the role of bacterial infection in the formation of these stones.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1442-2042.1996.tb00517.x
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  • 5
    Electronic Resource
    Electronic Resource
    Escherichia coli Virulence Factors and Serotypes in Acute Bacterial Prostatitis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    International journal of urology 4 (1997), S. 0 
    Details
    ISSN: 1442-2042
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Escherichia coli is the most frequent pathogen in both acute bacterial prostatitis and acute uncomplicated urinary infections. To assess the virulence profiles of E. coli in acute prostatitis, the serotypes and virulence factor (VF) genotypes were determined. Methods We studied 107 E. coli isolates from cases of acute bacterial prostatitis, 76 isolates from acute pyelonephritis, 194 isolates from acute cystitis and 80 fecal isolates from healthy people. All pyelonephritis and cystitis isolates were from women. Seven urovirulence determinants were analyzed by DNA colony hybridization, including the genes for type 1 fimbria (pil), P fimbria (pap), S fimbria (sfa), afimbrial adhesin AFA-I (afal), α-hemolysin (hly), cytotoxic necrotizing factor 1 (cnfl) and aerobactin (aer). 0:H:K serotypes were also determined. Results With the exception of pil and afal, all VFs were significantly more often associated with prostatitis, pyelonephritis and cystitis isolates than with the fecal isolates. The prevalence of sfa, hly and cnfl was higher in prostatitis isolates than in pyelonephritis and cystitis isolates, and the pap+sfa+hly+cnf+ genotype was dominant among prostatitis isolates (48.8%). Nine O serotypes(01, 02, 04, 06, 01 6, Ol 8, 022, 025 and 075) accounted for 79.4%, 73.7% and 78.4% of the prostatitis, pyelonephritis and cystitis strains, respectively. There was an apparent correlation between serotype and genotype in uropathogenic E. coli. Conclusion The predominance of O serotypes in female urinary tract infections and a high percentage of multiple VFs among the prostatitis isolates suggested that VFs play important roles in the pathogenesis of acute bacterial prostatitis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1442-2042.1997.tb00192.x
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  • 6
    Electronic Resource
    Electronic Resource
    Relationship Between Gastric Ulcer and Helicobacter pylori VacA Detected in Gastric Juice Using Bead-ELISA Method (2002)
    Oxford, UK : Blackwell Science Ltd
    Helicobacter 7 (2002), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background. VacA is an important pathogenetic factor produced by Helicobacter pylori. VacA has often been detected in supernatants of liquid cultures or lysates of whole bacterial cells. However, no studies have ever tried to assay VacA produced in the human stomach. We applied a very sensitive and simple method, bead-ELISA, to detect VacA in gastric juice.Materials and Methods. Forty-eight H. pylori-positive patients (16 nonulcer dyspepsia, 16 gastric ulcer, and 16 duodenal ulcer) and four H. pylori-negative nonulcer dyspepsia patients had endoscopy performed and gastric juice were aspirated. Polystyrene beads coated with the antibody to VacA, were used in this bead-ELISA method. The nucleotide sequences of vacA in the signal and middle regions were investigated.Results. Of the 48 samples that were positive for H. pylori, 21 [43.8%] were found to be VacA positive in gastric juice. The average and maximum concentrations of detected VacA in gastric juice were 143.2 ± 216.5 and 840 pg/ml, respectively. The average density of VacA from gastric ulcer patients (227.5 ± 276.7 pg/ml) was higher than that found in nonulcer dyspepsia (51.8 ± 39.8 pg/ml) and duodenal ulcer (49.2 ± 21.5 pg/ml) patients. There was no relationship between VacA in gastric juice and vacA genotype.Conclusions. VacA in gastric juice could be directly detected by bead-ELISA. In this study, the diversity of disease outcome was associated with not the quality but the quantity of VacA. Therefore, not only the quality but also the quantity of VacA is important etiological factors in the pathogenesis of mucosal damage.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.2002.00097.x
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  • 7
    Electronic Resource
    Electronic Resource
    Cloning of a Vero toxin (VT1, Shiga-like toxin I) gene from a VT1-converting phage isolated from Escherichia coli O157:H7 (1987)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 44 (1987), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Vero toxin (VT1, Shiga-like toxin I)-converting phages were induced with UV light from Escherichia coli O157:H7 strain 83–1386. A non-toxigenic E. coli C600 strain, lysogenized with a toxin-converting phage (86-02), produced VT1. A phage solution was prepared from the lysogenized E. coli C600 (86-02) strain and the phage DNA was prepared. EcoRI-fragments of the phage DNA were ligated with EcoRI-digested pBR325ΔTc and it was transformed into E. coli strain MC1061. Transformants with VT1 production commonly contained a 5.1-kb EcoRI-fragment. The restriction map of the EcoRI-fragment was prepared and it was found that a 2.1-kb BamHI-BglII fragment encoded VT1 production.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1987.tb02235.x
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  • 8
    Electronic Resource
    Electronic Resource
    Persistence of specific antigenic protein in the serum of chickens given intravenously botulinum toxin type B, C, D, E or F (1987)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 43 (1987), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A sublethal dose of Clostridium botulinum progenitor toxin of each of types B, C, D, E, and F was injected once intravenously into chickens. Blood samples were withdrawn periodically from the chickens to determine the toxin remaining in the serum by the mouse injection test and by enzyme-linked immunosorbent assay (ELISA) for both toxic and nontoxic components composing the progenitor toxin. Both components were detected by ELISA for at least a few days after the serum had became innocuous to mice, indicating a higher stability of the antigenicities of both components than the lethal toxicity in the chicken serum. For the diagnosis of botulism, it seems justified to recommend detection of the antigen (toxic component or nontoxic component or both) by ELISA even if no toxin is detected by the mouse test. Such immunological tests would no doubt contribute to an increase in the rate of diagnosis of human and animal botulism cases, particularly when blood sampling is delayed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1987.tb02172.x
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  • 9
    Electronic Resource
    Electronic Resource
    Distribution of the zot (zonula occludens toxin) gene among strains of Vibrio cholerae 01 and non-01 (1993)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 106 (1993), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract The distribution of the zot gene that encodes the zonula occludens toxin, a newly described toxin of Vibrio cholerae, among clinical, environmental and food isolates of V. cholerae 01 and non-01 was investigated. Both the zot gene and the ctx gene that encode cholera toxin were found in 247 of 257 clinical strains and 62 of 415 environmental or food isolates of V. cholerae 01. The zot gene, but not the ctx gene was found in 37 strains (one clinical strain and 36 environmental or food isolates). In addition, two of 31 clinical strains and six of 98 environmental or food isolates of V. cholerae non-01 possessed both the zot gene and the ctx gene. These results demonstrated the predominantly concurrent occurrence of the zot gene and ctx genes among strains of V. cholerae 01 which suggests a possible synergistic role of ZOT in the causation of acute dehydrating diarrhea produced by V. cholerae 01.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1993.tb05950.x
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  • 10
    Electronic Resource
    Electronic Resource
    Detection of urovirulence factors in Escherichia coli by multiplex polymerase chain reaction (1995)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 12 (1995), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Abstract Primers to amplify the genes encoding the virulence factors of uropathogenic Escherichia coli, such as pilus associated with pyelonephritis (pap), haemolysin (hly), aerobactin (aer) and cytotoxic necrotizing factor 1 (cnf1) genes, were designed. The above primers along with previously reported primers for S fimbriae (sfa) and afimbrial adhesin I (afaI) genes were combined to develop a multiplex polymerase chain reaction (PCR) for detection of the respective virulence factors and for the identification of uropathogenic E. coli. The multiplex PCR to detect pap, sfa, afaI, hly, aer and cnf1 genes was highly specific and the sensitivity was found to be about 5 × 103 colony forming units of E. coli per ml. A total of 194 E. coli strains isolated from patients with simple acute cystitis were examined by the multiplex PCR and the results were in complete agreement with that obtained by DNA colony hybridization test. The multiplex PCR developed was, therefore, concluded to be a useful, sensitive and rapid assay system to identify uropathogenic E. coli.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-695X.1995.tb00179.x
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