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A multiplex PCR system was developed for specific identification of genes encoding heat-labile (LTI) and heat-stable (STI and STII) toxins of enterotoxigenic Escherichia coli (ETEC) strains. In addition, primers specific for the E. coli gene coding for 16S rRNA were used as an internal control of the DNA amplification. The specificity of the method was validated by single PCR tests performed with...
Eighty isolates of Escherichia coli were collected in Northern Palestine throughout the 1996 to 2000 period from hospitalized patients with urinary tract infections (UTIs). Resistance rates were ampicillin, 65%; co-trimoxazole, 55%; cefuroxime, 10%; cefotaxime, 7.5%; ceftazidime, 2.5%; ciprofloxacin, 12.5%; gentamicin, 6.25% and amikacin, 1.25%. No imipenem-resistant isolates were identified. To determine...
Resistance to antituberculous agents is an important cause of ineffectiveness of antimicrobial therapy. The resistance of M. tuberculosis to antituberculous agents is a result of mutations in genes participating in those agent's action. The antituberculous drug - isoniazid can be activated by Mycobacterium tuberculosis either through a hydroperoxidase I/II or a superoxide-dependent oxyferrous pathway...
In this study, we evaluated three PCR-based methods for the molecular typing of nonpathogenic Fusarium oxysporum isolates: random amplified polymorphic DNA (RAPD), polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and amplified fragment length polymorphism (AFLP). The analyses were performed using 64 isolates of F. oxysporum collected from cotton-producing areas in Egypt...
To confirm the sensitivity of the polymerase chain reaction (PCR) technique (versus blood cultures) and to gain a better understanding of the incidence of true- and false-positive results when using this technique, one hundred randomly chosen patients treated operationally because of valve defects were examined. In our studies we found that PCR techniques using universal primers complementary to the...
Repetitive element polymorphism-PCR (REP-PCR) is one of the tools that has been used to elucidate genetic diversity of related microorganisms. Using the MB1 primer, REP-PCR fingerprints from 110 Bacillus strains within the "B.cereus group" have identified eighteen distinct categories, while other more distantly related bacterial species fell within six additional categories. All Bacillus...
Alternaría alternata were isolated and identified from root, foliage and soil of both wheat and rice crops and their aggressiveness was studied using aggressiveness analysis. Isolates of Alternaría alternata were genetically characterized using RAPD's. The investigations were based on surveys of wheat and rice crops in the rice-wheat cropping areas of Pakistan. The study showed that Alternaría alternata...
Extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae (EPKP) strains are frequently implicated in outbreaks in neonatal units. From April 2002 to January 2003, 149 neonates were colonized/infected with EPKP In the Neonatal Clinic of the Teaching Hospital at the Medical University of Gdansk, Poland. A novel assay based on suppression of PCR, ADSRRS-fingerprinting, was successfully evaluated...
PCR-RFLP analysis of commonly occurring insertion sequences ISS1-type, IS904 and IS982 in Lactococcus sp. and Leuconostoc sp. was used for the genetic differentiation of 17 strains of lactic acid bacteria. ISS1-type and IS982 were found in all analysed strains while 1S904 was present exclusively in strains belonging to Lactococcus sp. Amplification of ISS1-type IS sequences resulted in formation of...
Pulsed field gel electrophoresis (PFGE), multiplex PCR and multilocus sequence typing (MLST) methods were used for genotyping study of seventy-three L. monocytogenes isolates collected in Poland between 2000 and 2002 from human, food, environment and a diseased goat. The multiplex PCR, which is an alternative method to classical serotyping, divided the isolates into four PCR groups, IIa (42.5%), IIb...
Nine strains belonging to Bacillus cereus group has been isolated from food and environmental samples. Their taxonomic position was confirmed by RFLP analysis of 16S rRNA gene digested with Taq\. The detection of DNA sequences encoding the hemolysin BL complex and enterotoxin NHE, was studied in Bacillus sp. isolates. Set of primers was used to amplify fragment of hblD gene by PCR. For the detection...
The aim of this study was to evaluate methicillin resistance detection methods currently used when studying coagulase-negative staphylococci (CoNS). The resistance to oxacillin of 142 strains from seven species of CoNS isolated from the Intensive Care Unit environments was tested. The methods used were: disc diffusion test with cefoxitin (FOX₃₀) and oxacillin (OX₁), oxacillin agar screen test with...
In the present study the bacterial community structure in a sequencing batch reactor (SBR) operating in autotrophic conditions was determined in relation to temporal variations of NH₄-N, NO₂-N, NO₃-N levels in the effluent. Bacterial richness and composition were determined by PCR-DGGE (Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis) of 16S rRNA and amoA genes, and RISA (Ribosomal...
A number of Enterococcus strains with high-level inducible resistance to vancomycin have been identified, and the relative incidence of these strains has increased significantly in the last years. The first outbreak caused by vancomycin-resistant enterococci in Poland was reported in 1999. Vancomycin-resistant Enterococcus faecium is known for its propensity to cause infections which are difficult...
Wastewater treatment based on activated sludge is known to be one of the most effective and popular wastewater purification methods. An estimation of microbial community variability in activated sludge allows us to observe the correlation between a particular bacterial group's appearance and the effectiveness of the removal of chemical substances. This research is focused on microbial community temporal...
Multiplex-PCR assay for identification of Klebsiella pneumoniae isolates carrying gene clusters for biosynthesis of capsular polysaccharide (CPS) types K1 and K2 was developed. Genes wzc and orflO of the cps cluster were applied as K1 and K2 specific markers respectively. The assay specificity was confirmed using 147 isolates of Klebsiella spp. including 77 K-antigen reference strains. The multiplex-PCR...
In this study, we revealed rapid detection of streptomycin-producing Streptomyces spp. by extraction of total soil DNA from 14 soil samples using a modified lysis method followed by PCR amplification of a genus-specific sequence in the Streptomyces' 16S rDNA gene DNA band of the expected size (438 bp) was seen with all the samples. Additionally, specific amplification of the streptomycin-coding gene...
In search of an effective DNA typing techniaue for Acinetobacter baumannii strains for hospital epidemiology use, the performance and convenience of a new target seauence was evaluated. Using known genomic seauences of Acinetobacter baumannii strains AR 319754 and ATCC 17978, we developed single-locus variable-number direct-repeat analysis using polymerase chain reaction-restriction fragment length...
Two typing methods were evaluated, utilizing 62 clinical strains of Pseudomonas aeruginosa, to assess their usefulness as tools to study the bacterial diversity within this complex group. Genetic diversity was determined by PCR ribotyping and enterobacterial repetitive intergenic consensus (ERIC) PCR. By these methods, 9 and 36 genotypes were found, respectively. The result showed that ERIC PCR analysis...
The experiment was carried out in two SBR reactors differing in ammonia load (reactor R1 - ammonia load of 130 mg N-NH₄ x d⁻¹, reactor R2 - ammonia load of 250 mg N-NH₄ x d⁻¹). Feeding conditions in the reactors were switched from favoring autotrophic nitrification through favoring heterotrophic processes and back to autotrophic conditions. Observations of ammonia-oxidizing bacteria (AOB) community...
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