Significance and Impact of the Study: This study showed that serological analysis greatly complements bacterial isolation and helps in the diagnosis and confirmation of Shiga toxin (verotoxin)-producing Escherichia coli (STEC) infections. Serological tests can be performed to qualify the patient for the typical haemolytic uraemic syndrome (STEC-HUS). In Poland, STEC-HUS in children is mostly caused by the E. coli serotype O26, which indicates that there is an increasing number of non-O157 STEC infections associated with human diseases in Europe.
AbstractTypical haemolytic uraemic syndrome (STEC-HUS), caused by Shiga toxin (Stx)-producing Escherichia coli (STEC), is a serious, life-threating disease that mainly affects children. Bacteriological and genetic tests are commonly used in the routine laboratory diagnosis of STEC-HUS; however, serological methods have emerged as useful and reliable diagnostic tools, especially when bacterial isolation fails. In this study, we present the results of the serological investigation of 72 paediatric patients suspected for HUS, hospitalized during 2011-2019 at the Department of Pediatrics and Nephrology of Children's Hospitals in Poland. During the routine laboratory investigation STEC strains were isolated only from nine stool samples. However, serological investigations confirmed 45 cases of STEC infections in children with HUS. In this study, 22 (48Á9%) paediatric patients were infected by E. coli serotype O26, 11 (24Á4%) by serotype O145, 9 (20Á0%) by serotype O157, and 3 (6Á7%) by E. coli serotype O111. In the majority of these patients, in addition to a high level of IgA, IgG and IgM antibodies to lipopolysaccharide of particular E. coli serotypes, antibodies to recombinant proteins Tir, Stx2b and intimin were detected. Our results confirm that serological tests are useful in the diagnosis of STEC-HUS.
Introduction: Tularemia is a highly infectious zoonotic disease caused by Gram-negative bacterium Francisella tularensis. The microbiological diagnosis of tularemia is based mainly on serological investigations. The present study was undertaken to determine the avidity of IgG class antibodies to Francisella tularensis in the course of tularemia in humans and to evaluate its value for estimation of the phase of diseases.
Methods: Fifty two serum samples obtained from 40 patients with tularemia were tested by in-house ELISA in duplicate in the same plate, without and after the 0.5 h incubation with 8M urea. The age of the subjects was between 6 and 77 years. From one patient, a 9-years-old girl with oculoglandular form of tularemia, five serum samples were taken, respectively after 0.5, 1.5, 3, 6 and 12 months from the beginning of the first clinical symptoms.
Results: The results of the study showed higher values of the avidity index (AI) of IgG antibodies for F. tularensis, often exceeding the value of 0.9, in children and adolescents than in adults. The examination of serum samples obtained 2-3 times in the course of tularemia from few patients did not show significant differences in the level of avidity index depending on the period of the disease. However, in five serum samples obtained from a 9-years-old girl in the different phases of tularemia the avidity index showed increasing values (0.51, 0.80, 0.92, 0.90 and 0.94, respectively).
Conclusions: The avidity index of IgG may be helpful in excluding recent infection, but its usefulness in detecting an active phase of invasion requires further research.
Determination of immune status of patients to diphtheria toxin is based mainly on the results of commercially available ELISA kits. The aim of the present study was to compare the results obtained by ELISAs from seven different manufacturers: Mikrogen, Immunolab, Sekisui Virotech, NovaTec, Virion\Serion, IBL International and Euroimmun. All assays were performed according to the manufacturers' instructions. The concentrations of the anti-diphtheria toxin antibodies in 72 serum samples were calculated on the basis of curves constructed from standards supplied by manufacturers and the new reference material-International Standard for Diphtheria Antitoxin (10/262). The repeatability and reproducibility of all the ELISA kits tested were good. Number of sera with concentrations of the anti-diphtheria toxin antibodies below the WHO-recommended level of protection (0.1 IU/ml) were dependent on the ELISA used: Mikrogen, 20/72 samples (27.7%); Immunolab, 11/72 samples (15.3%); Sekisui Virotech, 0/72 samples (0%); NovaTec 18/72 samples (25.0%); Serion 12/72 samples (16.7%); IBL International, 7/72 samples (9.7 %); and Euroimmun, 17/72 samples (23.6%). However, the results obtained in particular ELISAs, with the exception of Sekisui Virotech, were much more consistent when the concentrations of the anti-diphtheria toxin antibodies in 72 sera measured by using curves constructed from the International Standard 10/262. The data obtained clearly demonstrated that manufacturer-dependent differences between anti-diphtheria IgG ELISA kits exist. The differences in recommendations accepted by the individual manufacturers together with differences shown in our studies in sensitivity greatly affect the clinical interpretation of results.
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