Biomedical subjects
Publications and source records attributed to H Chart.
Certain strains of enteroaggregative Escherichia coli cause detachment of HEp-2 cells during adhesion tests, preventing observation of the aggregative adherent phenotype. The use of formalin-fixed HEp-2 cells prevents cell detachment and facilitates the detection of enteroaggregative E. coli.
AIMS: To evaluate a rapid immunoblotting procedure for providing evidence of infection with Salmonella typhi using 73 sera from patients infected with S typhi. METHODS: A sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE)/immunoblotting procedure using lipopolysaccharide (LPS, O = 9,12) and flagellar (H = d) antigens was used. RESULTS: Seventy two of 73 sera contained antibodies to LPS, 40 sera also contained antibodies to H = d flagellar antigens. Analysis of acute and convalescent sera showed that only 62% of patients produced antibodies to flagellar antigens. CONCLUSIONS: The SDS-PAGE/immunoblotting procedure provided a rapid method for providing serological evidence of infection with S typhi.
A type of in vivo phenotype of Campylobacter jejuni was obtained by maintaining bacteria in the peritoneal cavities of chickens for one week. These bacteria, which had not been subcultured on laboratory media, were used to prepare outer membranes for comparison with C. jejuni grown in vitro. Flagella with subunits of 65 kDa and a single porin with a protein subunit of 49 kDa were expressed constitutively; however, outer membrane proteins of 55, 35 and 20 kDa, and intermediate-chain lipopolysaccharide were only expressed by bacteria maintained in chickens.
UNLABELLED: From September 1989 until September 1993, stool specimens and sera from 113 children with diarrhoea-associated haemolytic uraemic syndrome (HUS) from the Netherlands, two university hospitals in Belgium and one university hospital in Germany were examined for the presence of verocytotoxin-producing Escherichia coli (VTEC) infection. Evidence for VTEC infection was observed in 88 (78%) patients with HUS compared to 2 (3%) of the 65 children with acute gastro-enteritis Serotype O157 was the causative agent in 76 (86%) of these 88 patients with VTEC-associated HUS and verocytotoxin-2 (VT-2) was the most frequent toxin produced. Serological testing for antibodies to O157 O-antigen yielded the highest number of positive results compared to the other test methods. Antibodies to O157 were found in sera of 71 (65%) of 110 patients with HUS and one control serum. Stool and sera examination for VTEC in 95 family contacts of 28 patients with HUS demonstrated an evidence for VTEC infection 33 (35%). In contrast, in patients with HUS serological antibodies to O157 O-antigen were found in only 3 (4%) of 85 family contacts. CONCLUSION: In this part of Western Europe, VT2-producing Escherichia coli, mainly those belonging to serogroup O157, are the major cause of HUS in childhood.
Investigations were performed by the Laboratory of Enteric Pathogens on Vero cytotoxin-producing Escherichia coli (VTEC) in England and Wales from 1992-4. Bacterial isolates, faeces and sera obtained from patients with diarrhoea, bloody diarrhoea and haemolytic uraemic syndrome were examined. Using serotyping, Vero cytotoxin gene probing and serodiagnostic tests for E. coli O157, evidence of infection was detected in 543, 434 and 491 individuals in 1992, 1993 and 1994 respectively; VTEC of serogroup O157 were isolated from 470, 385 and 411 cases. The O157 VTEC strains belonged to at least 19 different phage types (PT) although 84% belonged to PT2, PT49, PT8, PT1 or PT4. Antibodies to E. coli O157 lipopolysaccharide were detected in 13% of the cases. The average annual rate of infection with O157 VTEC was 0.83/100000 and 12% of the 1458 individuals with evidence of infection with VTEC or E. coli O157 developed haemolytic uraemic syndrome. There were at least 18 general outbreaks and many family outbreaks.
Strains of Salmonella enteritidis, Salm. typhimurium and Salm. virchow, carrying different numbers of plasmids, were examined for the ability to multiply in sera. Viable counts were performed to monitor the kinetics of growth of bacteria when in human, chicken and turkey sera. The presence of plasmids in Salm. enteritidis, Salm. typhimurium and Salm. virchow reduced considerably the ability of strains of these serotypes to multiply in serum. SDS-PAGE was used to show that growth of Salm. enteritidis in serum did not involve changes in outer membrane proteins or lipopolysaccharide. It was concluded that the carriage of plasmids may be disadvantageous for the survival in serum of certain common salmonella serotypes.
The role of lipopolysaccharide (LPS) in the serotyping of Campylobacter jejuni based on heat-stable antigens was examined using SDS-PAGE and a silver stain for carbohydrate. None of the 32 type strains of Camp. jejuni expressed long-chain LPS. Rabbit antibodies, prepared to 10 selected strains of Camp. jejuni, reacted with surface-exposed carbohydrate antigens, which were not LPS. This study suggests that the heat-stable antigens of Camp. jejuni, which form the basis for the established Penner serotyping scheme, are probably capsular and not LPS.
One hundred sera from patients with haemolytic ureamic syndrome were screened for antibodies to the lipopolysaccharide (LPS) of Escherichia coli serogroup O5, O26, O115, O128, O145 and O153, and Shigella dysenteriae 1. Three sera contained antibodies to the LPS of E. coli belonging to serogroup O5.
Outer membrane-associated proteins of 18 kDa were expressed by enteroaggregative Escherichia coli (EAggEC) belonging to serotypes O126:H27 and O44:H18, which hybridized with a probe derived from a plasmid necessary for enteroaggregative adhesion. The 18 kDa proteins expressed by strains of E. coli, belonging to these serotypes, were surface exposed and antigenically similar but not structurally identical.
Using pulsed-field gel electrophoresis (PFGE) and 16S rRNA (rrn) analysis (ribotyping), the in vivo derivation of strains of Salmonella enteritidis PTs 9a and 7 from a strain of S. enteritidis PT 4 has been demonstrated. All strains were isolated from a single patient over a 6-week period. Further studies have demonstrated that in terms of pulsed-field profile and ribotype, the genotypes of the patient-derived strains differed from those of the reference strains of the respective phage types. It is concluded that when used in combination, these methods can provide evidence of phylogenetic relationships in apparently unrelated S. enteritidis phage types isolated during pathogenesis of disease.
Strains of Salmonella virchow express long-chain lipopolysaccharide which migrates as one of two distinct profiles during SDS-PAGE. Both LPS phenotypes were detected within a given strain of Salm. virchow and shared the O = 7 antigen.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
The pH of the environment influenced the expression of outer membrane proteins by S. enteritidis PT4 growing in broth. Growth in broth at pH 5 to 7 resulted in variation in expression of outer membrane proteins of 18 to 22 kDa. Bacteria became acid-fixed and non-viable following prolonged incubation in broth with a pH below 5, and expression of flagella was repressed.
Explore the source record for details and available documents.
Using pulsed-field gel electrophoresis (PFGE), nine profile types were identified in XbaI digests of a panel of strains of Salmonella enteritidis phage type 4 (PT 4) isolated in England and Wales between 1967 and 1992. When applied to 39 isolates from humans, chickens, poultry products and feed, strains belonging to pulsed-field profile (PFP) 1 predominated. DNA-DNA hybridization studies demonstrated that a band of approximately 57 kb was associated with the presence of the 38 MDa S. enteritidis 'serovar-specific' plasmid. It is concluded that PFGE provides a method for discriminating strains of S. enteritidis PT 4 suitable for epidemiological investigations.
Vero cytotoxin-producing Escherichia coli (VTEC) were isolated from the faecal specimen of a patient with haemolytic uraemic syndrome. The isolates belonged to two rare VTEC serotypes, O9ab:H- and O101:H-. Polymerase chain reaction gene amplification products were detected with primers specific for the VT2e gene, a variant of VT2. The toxin from both isolates was cytotoxic to Vero cells but not to HeLa cells. An 18 kb EcoRI restriction enzyme fragment of genomic DNA from both strains hybridised with a VT2 polynucleotide DNA probe.