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Biomedical subjects

S Tabaqchali

Publications and source records attributed to S Tabaqchali.

At least 73 records · Page 4Linked to original sources

The tangible cost implications of a hospital outbreak of multiply-resistant Salmonella.

A hospital outbreak of multiply-resistant Salmonella heidelberg infection, which affected 17 patients and 2 staff, is described. The tangible cost of the outbreak was estimated at 21 pounds 151, 17 pounds 989 (85.1%) of which was borne by the hospital. The cost to the Microbiology Department was 3596 pounds (17.0% of the total). A detailed analysis of the costs and implications for staffing disruption is given and a comparison is made with the costs of preventive activities. Ways of containing expenses in the event of an outbreak and the economic implications for clinical budgeting and privatization of the laboratory service are considered.

Clinical Laboratory Techniques↗

Molecular cloning and expression of Campylobacter pylori species-specific antigens in Escherichia coli K-12.

A gene bank of Campylobacter pylori DNA in Escherichia coli was constructed by cloning Sau3A-cleaved DNA fragments into the bacteriophage vector lambda EMBL3. The expression of C. pylori antigens was determined by screening the gene library with adsorbed C. pylori whole-cell rabbit antisera. One recombinant clone which reacted positively (lambda CP2) was studied further. Immunoblot analysis with lambda CP2 showed a polypeptide band of 66 kilodaltons (kDa) reacting antigenically with the adsorbed antiserum. Extraction of DNA from lambda CP2 and digestion with SalI revealed a DNA insert of 17 kilobases (kb). Subcloning with SalI and the E. coli vector pUC18 showed that the DNA also encoded a 31-kDa antigen. The cloned antigens were shown by immunoblotting to have the same molecular weight in E. coli as in C. pylori and to be present in all C. pylori strains. Antiserum was raised against the cloned polypeptides and found to react only with C. pylori when analyzed by dot blotting and indirect immunofluorescence. The cloned antigens were determined to be expressed from the pUC18 lac promoter. The DNA encoding these antigens was radiolabeled with 32P and found to hybridize only to C. pylori strains. Immunoblotting with affinity-purified polyclonal antibody to the urease enzyme of C. pylori revealed that the cloned antigens may be part of the urease enzyme.

Antigen-Antibody Reactions↗

Histamine synthesis by respiratory tract micro-organisms: possible role in pathogenicity.

Five bacterial species considered to be potential pathogens in acute exacerbations of chronic bronchitis, cystic fibrosis, and pneumonia--Branhamella catarrhalis, Haemophilus parainfluenzae, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus pneumoniae--were evaluated for their potential to synthesise histamine in vitro. Bacterial species commonly isolated from infected sputum but generally not considered to be pathogenic--Enterobacteriacae, Neisseria pharyngis, coagulase negative staphylococci, alpha-haemolytic streptococci, and Candida albicans--were similarly studied. Of the "pathogens", the Gram negative species B catarrhalis, H parainfluenzae and Ps aeruginosa synthesised clinically important amounts of histamine; this was not the case for the Gram positive species S aureus and S pneumoniae. Of the "non-pathogenic" species, only the Enterobacteriacae, as a group, were found to synthesise clinically important amounts of histamine. These results show that some Gram negative bacteria, associated with acute exacerbations in respiratory infections, produce histamine and possibly other inflammatory mediators, which may contribute to their pathogenecity in the lower respiratory tract in vivo.

Bacteria↗

Epidemiology of Aeromonas species in a hospital.

The occurrence of Aeromonas spp. in a hospital water supply, in patients and in the local community was investigated. In healthy persons outside the hospital the isolation rate was 3.6% and among hospital patients it was 6%. Seven per cent of water samples yielded Aeromonas strains. Isolates were typed by sodium dodecyl sulphate polyacrylamide gel electrophoresis of sulphur-35 methionine labelled proteins of Aeromonas isolates. No relationship between water and human isolates could be established, even when a strain of A. hydrophila producing Vero cell cytotoxin contaminated enteral feeds given to patients in the intensive care unit.

Aeromonas↗

Clostridium difficile infection in patients with haematological malignant disease. Risk factors, faecal toxins and pathogenic strains.

Two hundred and forty-eight patients from shared oncology and general medical wards were prospectively studied over a 6-month period for carriage of Clostridium difficile during an outbreak of clinical disease with an epidemic strain of the organism. Risk factors for infection were assessed. Acute leukaemia and/or its treatment were identified as significantly increasing the risk of infection. The relationship between the type of C. difficile isolated (as defined by a typing system based on the incorporation of [35S]methionine into bacterial proteins followed by gel electrophoresis), the presence of faecal toxins A and B and clinical symptoms were analysed. Carriage of the epidemic strain, type X, had a significant association with symptoms amongst oncology patients, with two thirds of these patients having detectable faecal toxin A and one third detectable faecal toxin B. During an outbreak of C. difficile-associated disease, typing the organism and assaying for both faecal toxins in symptomatic patients may be of benefit in determining which patients require specific, urgent treatment.

Adolescent↗

Identification of gram-positive anaerobic cocci by use of systems for detecting pre-formed enzymes.

Two systems for detecting pre-formed enzymes, RapID ANA and a prototype system from API, were compared in a blind study for their ability to identify 69 gram-positive anaerobic cocci isolated from clinical specimens. Both systems were able to identify Peptostreptococcus anaerobius, Ps. asaccharolyticus and Ps. micros accurately without the need for further tests. The prototype API system identified all isolates of Ps. magnus correctly, but the RapId ANA system misidentified several isolates as Ps. micros. Numerous different enzyme patterns were found with the indole-negative, butyrate-producing cocci (Ps. prevotii and Ps. tetradius), suggesting that this group of organisms may be heterogeneous. We conclude that kits for detecting preformed enzymes are of considerable potential for the identification of gram-positive anaerobic cocci in clinical laboratories.

Enzymes↗

Preformed enzyme profiles of reference strains of gram-positive anaerobic cocci.

The preformed (constitutive) enzyme profiles of 30 type strains and reference strains of gram-positive anaerobic cocci were determined with two commercial systems, RapID ANA and a prototype system from API. Both systems identified Peptostreptococcus anaerobius, Ps. asaccharolyticus, Ps. indolicus, Ps. magnus and Ps. micros accurately, except for one strain of Ps. magnus misidentified as Ps. micros by the RapID ANA system. The indole-negative, butyrate-producing cocci (classified at present as Ps. prevotii and Ps. tetradius) produced several different, unique patterns with the prototype API system, but the results with RapID ANA were often misleading. Eight strains of Hare group cocci produced previously described profiles. Four strains of streptococci produced profiles easily distinguished from those of the gram-positive anaerobic cocci. We conclude that most gram-positive anaerobic cocci can be identified rapidly and reliably to the species level by their preformed enzyme profiles, providing that their underlying classification is sound. Problems were encountered with the butyrate-producing cocci, which appear to be a more heterogeneous group of organisms than is currently acknowledged; further taxonomic studies on these organisms are required.

Animals↗

Identification and molecular cloning of a 70 kDa species-specific antigen common to Clostridium difficile.

Three common antigens (CB 1, 2 and 3), characteristic of Clostridium difficile species were identified by immunoblot analysis using homologous and heterologous rabbit antisera, raised against whole cells from 9 distinct strains of C. difficile. A gene library of C. difficile genomic DNA was constructed in Escherichia coli by cloning in Sau 3A-cleaved clostridial DNA fragments into the bacteriophage vector lambda EMBL3. OUt of 3000 plaques screened using the whole cell antisera, 27 clones were positively identified. One of these clones, designated gamma Cd21, expressed high levels of an antigen which could be immunologically identified using whole cell antisera against the 9 C. difficile strains. Antiserum raised against the clone gamma Cd21 identified a 70 kDa antigen (previously named CB1) as demonstrated by immunoblot analysis. Monospecific antiserum against gamma Cd21 recognises the 70 kDa antigen in all 97 strains of C. difficile derived from worldwide sources and does not cross-react with 17 strains from 13 other clostridial species.

Antigens, Bacterial↗

Molecular cloning and genetic analysis of a chloramphenicol acetyltransferase determinant from Clostridium difficile.

A gene bank from a clinical isolate of Clostridium difficile expressing high chloramphenicol acetyltransferase activity was constructed by cloning Sau3A-cleaved clostridial DNA fragments into the plasmid vector pUC13. Among 1,020 clones tested, 11 were resistant to chloramphenicol; 1 of these, with an insert size of 1.9 kilobases (pPPM9), was studied further. The clone pPPM9 was mapped using a variety of restriction enzymes, and a 0.27-kilobase EcoRV-TaqI restriction fragment was shown to be within the chloramphenicol resistance (Cmr) gene by using transposon (Tn1000) mutagenesis. The 0.27-kilobase fragment and the 1.9-kilobase insert were radiolabeled and used as DNA probes in hybridization studies. Southern blot analysis with the gene probes against chromosomal DNA from Cmr strains of C. difficile obtained from five distinct geographical locations revealed that at least two copies of the same chloramphenicol acetyltransferase gene were present for each strain. Hybridization of the gene probes against Cmr strains of Staphylococcus epidermidis, Staphylococcus aureus, Klebsiella edwardsii, Escherichia coli, and to four other clostridial species revealed no homology even under conditions of low stringency.

Blotting, Southern↗

Clostridium difficile and acute enterocolitis.

Clostridium difficile belonging to groups not normally detected in infancy was the only potential pathogen detected in the stools of two infants with severe enterocolitis. Further information regarding the virulence of this organism was obtained by use of a recently introduced typing scheme.

Clostridium↗

Anaerobic infections in the head and neck region.

Anaerobic bacteria form the predominant flora of the oral cavity, outnumbering facultative organisms by 10-1,000: 1. The type of anaerobic bacteria and their concentration depend on the anatomical site and the degree of anaerobiosis in the different sites in the mouth. Three groups of anaerobic bacteria inhabit the oral cavity; the strict anaerobes, the moderate anaerobes, and the microaerophilic group of organisms. The majority of anaerobic bacterial infections occurring in the region of the mouth, head and neck are caused by the commensal flora. These infections include dental and periodontal disease where the predominant organisms are Bacteroides species, Veillonella, Bifidobacteria, Peptococcus, Peptostreptococcus and Propionibacterium species. More recently, Bacteroides endontalis has been isolated from a periapical abscess of endodontal origin and B. gingivalis, B. intermedius, Haemophilus actinomycetemcomitans and Wollinella species in chronic periodontal disease. Treponema species and other strict anaerobes are seen in smears of severe periodontal disease and acute necrotising gingivitis, but have not yet been isolated in pure culture. Until such time, their role in disease remains uncertain. Fusobacterium nucleatum is specially associated with severe orofacial infections which may extend into the mediastinum. Other anaerobic infections include chronic otitis media, chronic sinusitis and mastoiditis, and brain abscess. Treatment of these conditions should include the use of beta-lactamase resistant antimicrobials, such as clindamycin or one of the nitroimidazoles with penicillin.

Abscess↗

Molecular cloning and expression of Clostridium difficile toxin A in Escherichia coli K12.

Clostridium difficile toxin A was purified to homogeneity and was used to raise monospecific antiserum in rabbits. A gene bank of C. difficile DNA in Escherichia coli was constructed by cloning Sau3A-cleaved clostridial DNA fragments into the bacteriophage vector lambda EMBL3. Out of 4500 plaques screened with antitoxin A, 9 clones were positively identified. One of these clones lambda tA5 expressed a 235 kDa protein which exhibited a cytotonic effect on Chinese hamster ovary cells, and had the ability to haemagglutinate rabbit erythrocytes, both properties characteristic of toxin A. The size of the lambda tA5 insert DNA was 14.3 kb.

Animals↗

Immunochemical characterization of a polysaccharide antigen of Bacteroides fragilis with an IgM monoclonal antibody.

An IgM mouse monoclonal antibody (McAb) Bf4 was produced to a surface polysaccharide of Bacteroides fragilis NCTC 9343. Immunoblotting showed that McAb Bf4 reacted strongly with a high molecular mass structure which was sensitive to oxidation with periodate but resisted protease treatment. An inhibition enzyme-linked immunosorbent assay (ELISA) indicated that McAb Bf4 did not cross react with the sixteen Bacteroides species and strains tested. Cells of B. fragilis NCTC 9343 recovered from the various interfaces of a Percoll discontinuous density gradient were tested in the inhibition ELISA. Bacteria from the 0-20%, 20-40% and 40-60% interfaces inhibited the ELISA; however, cells from the 60-80% interface did not. Electron microscopy with immunogold labelling showed that McAb Bf4 did not react with the extracellular fibrous network on bacteria recovered from the 0-20% interface, or the extracellular electron dense layer on cells from the 60-80% interface; however, it was associated with a surface structure on cells from the 20-40% interface. Growth in vivo did not enrich for bacteria with this structure.

Antibodies, Monoclonal↗

Typing of Aeromonas species by polyacrylamide-gel electrophoresis of radiolabelled cell proteins.

A method for typing species of Aeromonas by 35S-labelling and sodium-dodecyl sulphate polyacrylamide-gel electrophoresis (SDS-PAGE) of total soluble proteins is described. Radiolabelled-protein profiles from different strains were compared by use of the Dice coefficient. Typability was 100% and discrimination of pairs of differing strains was satisfactory with acceptable reproducibility. The results also yielded information on subgroups of phenotypic species.

Aeromonas↗

Quantitative bacteriology of the vaginal flora during the menstrual cycle.

The vaginal bacteriology of 10 healthy asymptomatic women was assessed during the menstrual cycle. Samples were taken from the posterior vaginal fornix for quantitative analysis. There were no significant alterations in the total vaginal flora at different stages of the menstrual cycle. The mean number of species isolated per specimen declined from 4.6 in week 1 to 2.9 in week 4. This decline was not caused by a decrease in the occurrence or concentration of any one organism or group of organisms. The vaginal pH decreased from a mean of 6.6 in week 1 to 4.3 in week 4, this increased acidity could not be attributed to the action of lactobacilli as their total incidence or concentration did not change during the menstrual cycle.

Bacteria, Aerobic↗