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Biomedical subjects

H Chart

Publications and source records attributed to H Chart.

At least 19 recordsLinked to original sources

Human serum antibody response to Helicobacter pylori whole cell antigen in an institutionalized Bangladeshi population.

AIMS: To use a commercial ELISA kit and an immunoblot assay to investigate the antibody levels of selected members of the Bangladeshi population to Helicobacter pylori protein antigens. METHODS AND RESULTS: Using immunoblotting, high seroprevalence rates were observed in all age groups, although the subjects within the 1-9 years age group had the highest seroprevalence of antibodies to H. pylori antigens. By ELISA, the highest level of seroprevalence was observed in those over the age of 20 years. CONCLUSION: On the basis of these results the overall prevalence rate of H. pylori infection for the whole population was 77.4%; 77.9% for orphan boys and 76% for carers. CagA antibodies were detected in 86% of those with high levels of antibodies to H. pylori antigens. SIGNIFICANCE AND IMPACT OF THE STUDY: A combination of immunoblotting and ELISA was the most efficient means of detecting serum antibodies to H. pylori antigens and could be applied to the screening of human sera for H. pylori-specific antibodies.

Adolescent↗

An investigation into the pathogenic properties of Escherichia coli strains BLR, BL21, DH5alpha and EQ1.

AIMS: To examine Escherichia coli strains EQ1, DH5alpha, BLR and BL21 for known pathogenic mechanisms. METHODS AND RESULTS: Using specific DNA probes, the strains were shown not to carry the genes encoding invasion, various adhesion phenotypes or expression of a range of enterotoxins. The strains were unable to express long-chain lipopolysaccharide and were susceptible to the effects of serum complement. Using a BALB/c mouse model, the strains were shown to be unable to survive in selected tissues or to persist in the mouse gut. Using a chick model, strains EQ1, BLR and BL21 invaded livers but not spleens; only strain EQ1 persisted in the chick gut. In Merino sheep, only strain EQ1 was detected 6 d post-infection. CONCLUSIONS: Escherichia coli strains EQ1, DH5alpha, BLR and BL21 did not carry the well-recognized pathogenic mechanisms required by strains of E. coli causing the majority of enteric infections. SIGNIFICANCE AND IMPACT OF THE STUDY: Escherichia coli strains EQ1, DH5alpha, BLR and BL21 were considered to be non-pathogenic and unlikely to survive in host tissues and cause disease.

Animals↗

The serodiagnosis of infection with Salmonella typhi.

BACKGROUND/AIMS: The serodiagnosis of infection with Salmonella typhi, using the Widal agglutination assay, relies on patients' antibodies to the O = 9,12 lipopolysaccharide (LPS) antigens, H = d flagellar antigens, and the Vi capsular antigens. A Vi agglutination titre of > 1/40 has traditionally been regarded as indicative of recent infection with S typhi. In this study, 91 sera were used to assess the reliability of the Widal agglutination assay based on antibodies to the Vi antigens. METHODS: The Widal agglutination assay was carried out using protocols established by the Central Public Health Laboratory, Colindale. Antibodies to the Vi capsular antigen were detected using a standard preparation of S typhi, ViI Bhatnagar variant strain (S typhi, ViI). Sera used in the study comprised 73 from patients who were culture positive for S typhi, 10 from patients who were culture positive for other species of Salmonella not expressing a Vi antigen (namely, S javiana, S enteritidis, S typhimurium, S stanley, S saint paul, S bareilly, or S mbandaka), and eight from healthy blood donors. RESULTS: Agglutination titres of > or = 1/40 were detected to S typhi ViI in 69 of 73 sera from patients with typhoid, although 27 of these also agglutinated an unrelated control antigen. The Widal assay also detected significant amounts of agglutinating antibodies to S. typhi ViI in all eight control sera and seven sera from patients infected with S bareilly, S enteritidis, S javiana, S mbandaka, S saint paul, and S stanley. CONCLUSIONS: Agglutinating antibodies to the Vi antigen can be detected by the Widal assay, but even with the appropriate control antigens the results were unreliable. The serodiagnosis of infections with S typhi should be based on the detection of antibodies to both the O = 9,12 LPS antigen and the H = d flagellar antigen by immunoblotting, and should not use the Vi antigen-based Widal assay. Conclusions should be made in the light of patients' clinical details and any knowledge of previous immunisation for typhoid.

Agglutination Tests↗

Campylobacter coli strains with enlarged flagellin genes isolated from river water.

A group of campylobacters isolated from river water were found to possess unusually large flagellin genes. Both phenotype and serology were consistent with identification as Campylobacter coli. Phylogenetic analysis of small (16S, rrs) and large subunit (23S, rrl) rRNA genes of a representative strain, NCTC 13006, demonstrated high levels of relatedness with C. jejuni and C. coli (99.1 and 98.3% similarity for 16S; 99.3 and 99.4% similarity for 23S). Large flagellin proteins were demonstrated by SDS-PAGE analysis. The flaA and flaB genes were sequenced and aligned with known campylobacter flagellin amino acid sequences. The encoded FlaA protein of the new group exhibited a high degree of divergence from other Campylobacter species. Within the central variable region of FlaA, a further hypervariable domain was identified containing characteristic repeated motifs. Separate pairwise alignments performed for the variable regions of the polypeptide indicated these large fla genes were more closely related to those of C. upsaliensis than to those of C. coli or C. jejuni.

Amino Acid Sequence↗

Characterization of enteroaggregative Escherichia coli isolated from outbreaks of diarrhoeal disease in England.

Twenty-two strains of enteroaggregative Escherichia coli (EAggEC), isolated from four outbreaks of diarrhoeal disease in England, were examined for a range of phenotypic attributes including the ability to produce fimbriae, haemolysins and siderophores, and cell-surface properties such as surface charge and hydrophobicity. Strains of EAggEC isolated from two of these outbreaks belonged to a diverse range of serotypes and were heterogeneous in phenotype. Strains of EAggEC isolated from the other two outbreaks belonged predominantly to serotypes 086:H34 and 098:H-, respectively. Only two strains expressed fimbriae and two strains produced an 18 kDa membrane associated protein (MAP), suggesting that EAggEC express a range of adhesion mechanisms to produce the cell arrangement recognized as the 'stacked brick' formation. The possible explanation for the diversity of EAggEC serotypes is discussed.

Bacterial Outer Membrane Proteins↗

Serodiagnosis of infection with verocytotoxin-producing Escherichia coli.

Human sera (167) were screened for antibodies to lipopolysaccharide (LPS) prepared from strains of Verocytotoxin-producing Escherichia coli (VTEC) belonging to a range of serogroups, secreted proteins expressed by attaching and effacing VTEC, enterohaemolysin and H = 7 flagellar proteins. Twelve sera (about 7%) contained antibodies to the LPS of E. coli 05 (one), 026 (two), 0115 (two), 0145 (one), 0163 (one) and 0165 (five). Sera containing antibodies to the LPS of E. coli O26 and O145 also contained antibodies to secreted proteins of 100 and 40 kDa. An additional 34 sera, known to contain antibodies to the lipopolysaccharide of E. coli O157, were examined for antibodies to enterohaemolysin, H = 7 flagellar antigens and bacterial cell surface-associated proteins of 5, 6 and 22 kDa. Three sera contained antibodies to enterohaemolysin and one serum contained antibodies to flagellar proteins. Antibodies to membrane-associated proteins were not detected. It was concluded that enterohaemolysin, H = 7 flagellar proteins and the cell surface-associated proteins were unsuitable for use in immunoassays for providing evidence of infection with VTEC.

Adolescent↗

The serodiagnosis of infections caused by Verocytotoxin-producing Escherichia coli.

Patients with haemolytic uraemic syndrome (HUS) and haemorrhagic colitis (HC) produce serum antibodies to the lipopolysaccharides (LPS) of Escherichia coli O157 and certain other E. coli serogroups. Patients may also make salivary antibodies to the LPS of E. coli O157. Serological tests based on these antibodies can be used to provide evidence of infection in the absence of culturable VTEC or the toxins they produce. Serum antibodies to LPS persist for several months following onset of disease, enabling both current and retrospective serological testing. The LPS of E. coli O157 shares epitopes with strains of Brucella abortus, Yersinia enterocolitica O9, Vibrio cholerae O1 Inaba, group N Salmonella and certain strains of Citrobacter freundii and E. hermanni. Serological tests for serum antibodies to E. coli O157 should be evaluated in the light of these cross-reactions. Serological tests to supply evidence of infection with E. coli O157 have been shown to provide a valuable adjunct to bacteriological procedures for detecting culturable VTEC and VT. The use of well characterized LPS antigens in association with the techniques of ELISA and immunoblotting provide valuable procedures for detecting evidence of infection with E. coli O157 and possibly other VTEC.

Antibodies, Bacterial↗

Evaluation of a latex agglutination kit for the detection of human antibodies to the lipopolysaccharide of Escherichia coli O157, following infection with verocytotoxin-producing E. coli O157.

A total of 150 human sera was used to evaluate a commercial latex agglutination kit for detecting antibodies to the lipopolysaccharide of Escherichia coli O157. A comparison of the kit with SDS-PAGE and immunoblotting showed that the kit had a sensitivity of 94.12%, a specificity of 99.15%, a positive predictive value of 96.97% and a negative predictive value of 99.15%.

Adolescent↗

Verocytotoxin-producing Escherichia coli infection in household members of children with hemolytic-uremic syndrome in The Netherlands.

BACKGROUND: Strains of verocytotoxin-producing Escherichia coli (VTEC) belonging to serogroup O157 (O157 VTEC) can cause a spectrum of disease that includes nonspecific diarrhea, hemorrhagic colitis and the diarrhea-associated form of the hemolytic uremic syndrome (D+ HUS). METHODS: We conducted a retrospective study of 34 children with D+ HUS caused by O157 VTEC to determine the frequency of VTEC infection in their household members. RESULTS: Gastrointestinal tract symptoms were reported in 1 or more household contacts of 17 (50%) of the 34 index cases. Of the 26 household members with gastrointestinal tract symptoms, 15 were parents and 11 were siblings. Evidence of VTEC infection was reported in 1 or more household contacts in 23 (68%) of the 34 families (in 46% of the siblings and in 28% of the parents). Nineteen (48%) siblings had a positive stool sample and in only 5 (12%) of the siblings IgM class serum antibodies to O157-lipopolysaccharide (LPS) were detected. Nineteen (31%) parents had a positive stool sample. Antibodies to O157-LPS were not detected in any of the parents. The occurrence of (bloody) diarrhea significantly correlated with the occurrence of IgM class serum antibodies to O157-LPS. CONCLUSIONS: It was concluded that household members of children with D+ HUS are often asymptomatically infected with O157 VTEC. Differences in the pathogenesis of the infection between infected individuals may be related to differences in the number of ingested O157 VTEC bacteria and to differences in susceptibility.

Antibodies, Bacterial↗

Growth of Salmonella enteritidis and S. pullorum on Hektoen agar and the expression of lipopolysaccharide or flagella.

Growth of strains of Salmonella enteritidis and Salmonella pullorum on Hektoen agar has been reported to influence the expression of long-chain lipopolysaccharide and motility respectively. In this study we used a panel of strains of S. enteritidis and S. pullorum to investigate these phenomena. Culture on Hektoen agar did not cause rough strains of S. enteritidis to express long-chain lipopolysaccharide or strains of S. pullorum to become motile. It was concluded that growth of strains of S. enteritidis and S. pullorum on Hektoen agar would not normally affect the expression of somatic or flagellar antigens, and would not influence the interpretation of the Kauffman-White typing scheme.

Agar↗

Expression of membrane-associated proteins by strains of enteroaggregative Escherichia coli.

Certain strains of enteroaggregative Escherichia coli express an outer membrane-associated protein, involved with the adhesion of these bacteria to HEp-2 cells. Strains of enteroaggregative E. coli hybridising with DNA probes for aggregative adhesion, diffuse adhesion and aggregative adhesion fimbriac II expressed an outer membrane-associated protein of 18 kDa regulated by magnesium ions. Strains hybridising with the aggregative adhesion probe only expressed a 20-kDa outer membrane-associated protein regulated by calcium and magnesium. The present study describes two populations of enteroaggregative E. coli which appear to adhere to HEp-2 cells by expressing antigenically distinct, negatively charged membrane-associated proteins.

Animals↗

Serum antibodies to secreted proteins in patients infected with Escherichia coli O157 and other VTEC.

Certain strains of verotoxigenic Escherichia coli (VTEC), and in particular those belonging to serogroup O157, cause attaching and effacing (AE) lesions of the host gut mucosa during pathogenesis. The mechanisms involved with bacterial attachment and the destruction of microvilli are determined by a cluster of genes within the LEE region, which also encode five secreted proteins. Sera from patients with antibodies to the lipopolysaccharide (LPS) of E. coli O157 and other VTEC were tested for antibodies to these secreted proteins. Twenty-one of 34 (62%) sera with antibodies to the lipopolysaccharide (LPS) of E. coli O157 also contained antibodies to one or more of the secreted proteins. Five of 12 sera containing antibodies to the LPS of a range of other VTEC serogroups also contained antibodies to 1 or more of the 5 secreted proteins, as did 16 of 70 (23%) sera from patients with haemolytic uraemic syndrome (HUS), haemorrhagic colitis (HC) or diarrhoea, but without bacteriological evidence of infection with VTEC and which did not contain antibodies to VTEC serogroups O5, O115, O145, O153 or O157. The detection of serum antibodies to secreted proteins may provide additional information for interpreting the results of established lipopolysaccharide-based VTEC serology.

Antibodies, Bacterial↗

Enteroaggregative Escherichia coli.

Enteroaggregative Escherichia coli (EAggEC) are a group of E. coli characterized by the ability to adhere to cultured cell monolayers with an aggregative or 'stacked brick' adhesion phenotype. These strains of E. coli are distinct from other pathogenic groups of E. coli. Epidemiological evidence suggests that strains of EAggEC are a significant cause of protracted diarrhoea in children, and may cause diarrhoea in adults. The group is heterogeneous, comprising a diverse range of serotypes that possess a variety of putative virulence factors. These include an enterotoxin similar to the heat-stable enterotoxin of enterotoxigenic E. coli, putative haemolysins and toxins, and various types of fimbriae and outer membrane proteins that may be involved in the adhesion process. The role of these factors in the production of disease is unclear. Diagnosis of EAggEC infection is problematic; serotyping alone cannot identify strains of E. coli expressing an EAggEC phenotype. Currently, EAggEC are identified by either cell adhesion tests or DNA-based tests involving gene probes or polymerase chain reaction (PCR) for detecting the genes encoding cell adhesion. The nature and significance of strains of E. coli expressing an EAggEC phenotype are poorly understood. This article reviews the current literature and speculates on the direction of future studies to define this emerging group of bacteria.

Adult↗