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Biomedical subjects

D G Fleck

Publications and source records attributed to D G Fleck.

At least 19 recordsLinked to original sources

Direct agglutination test and other assays for measuring antibodies to Toxoplasma gondii.

The performance of a direct agglutination test for the detection of toxoplasma specific IgG immunoglobulin was compared with that of the latex agglutination test. The direct agglutination test was less sensitive but more specific than the latex agglutination test. Quantitative results were not directly comparable, reflecting the different antibody profiles detected in each assay. The direct agglutination test represents an alternative to the latex agglutination test as a screening test for toxoplasmosis. Patients at risk of life threatening infection require detailed serological examination using additional assays.

Agglutination Tests↗

Five commercial enzyme linked immunosorbent assay kits for toxoplasma specific IgM antibody.

Five commercially available enzyme linked immunosorbent assay (ELISA) kits for the detection of specific IgM against Toxoplasma gondii were evaluated in a three centre study and results compared with those of the Public Health Laboratory Service ELISA for Toxoplasma IgM (PHL IgM ELISA). Fifty selected sera were tested by all the methods (Toxo-M, Captia Toxo-M EIA, Toxo Enz M EIA, Toxonostika IgM EIA, Sopazyme Toxo IgM EIA) at the three reference centres in England and Wales and 177 routine sera by all the methods in one or other of the centres. Ten of the 50 selected sera contained autoimmune antibodies but no specific IgM and 29 had toxoplasma specific IgM detectable by the PHL IgM ELISA. The kits were assessed for their specificity and sensitivity compared with the PHL IgM ELISA, and the percentage coefficient of variation for binding to the solid phases was determined. They were also rated subjectively by the staff performing the assays and an overall impression of each kit was gained by allocating scores of several criteria. There was quite close agreement among the results obtained with all five commercial assays and the PHL IgM ELISA, although some of the sera pre-selected as being potentially problematic showed the limitations of some of the assays.

Animals↗

Public Health Laboratory Service enzyme linked immunosorbent assay for detecting Toxoplasma specific IgM antibody.

An enzyme linked immunosorbent assay (ELISA) based on the antibody class capture method for the detection of specific IgM against Toxoplasma gondii, using the microtitre plate format, was developed. Antigen binding was detected using a monoclonal antibody, CIE3, conjugated to horseradish peroxidase. Prior mixing of the conjugate and antigen improved the stability of these reagents as well as removing an incubation stage from the assay. The incubation time of less than four hours permits a rapid throughput of specimens. Using the assay, a total of 163 sera were examined in a three centre study and good agreement was found. Results were expressed as arbitrary enzyme immunoassay units (EIUs) against a freeze dried standard. Throughout the study the standard serum showed a coefficient of variation less than 10% across the microtitre plate. By measuring IgM titres in patients having toxoplasmic lymphadenopathy with a known date of onset, IgM class antibodies were shown to peak at two months, persisting for about six months. In addition, a case of laboratory acquired toxoplasmosis was monitored. Sera shown to contain rheumatoid factor and antinuclear factor did not give false positive results. This rapid, robust, and simplified assay is used by the Public Health Laboratory Service Toxoplasma Reference Units and will provide a standard with which other assays can be compared.

Animals↗

Disseminated toxoplasmosis in cardiac transplantation.

The first case of disseminated toxoplasmosis following cardiac transplantation in the UK is described, with details of Toxoplasma antibody tests made on other cardiac transplant patients. Sixteen of 40 (39%) of recipients had Toxoplasma antibody before operation. Eleven of 30 (37%) of donors had Toxoplasma antibody. The were four occasions when a negative recipient received a heart from a positive donor. Three survived the immediate postoperative period and two became infected with toxoplasmosis. The implications of this are discussed. Disseminated toxoplasmosis appears much more often when heart muscle from a dye test positive donor is given to a dye test negative recipient. Antibiotic therapy is limited by the fact that the antitoxoplasma drugs available are static in their effect, and need to be given for prolonged periods postoperatively.

Adult↗

Comparative study of three tests (dye test, indirect haemagglutination test, latex agglutination test) for the detection of antibodies to Toxoplasma gondii in human sera.

An evaluation has been made of a commercial latex agglutination test, Toxotest-MT (TMT) (Eiken, Japan), for the detection of antibodies to Toxoplasma gondii in human sera. In qualitative studies, 878 sera were examined in both the TMT and dye test (DT) and 96.6% agreement was found, In quantitative studies 339 sera were titrated in the TMT and DT, with 337 of these sera also titrated in the indirect haemagglutination test (IHAT). Agreement between the DT and TMT was best, 78% of the sera showing titres within +/- 1 dilution in the two tests. The IHAT and TMT gave 66% agreement, while the DT and IHAT showed least agreement, 40.9%. The results suggest that the pattern of antigenic determinants to which antibody levels are measured are different in the three test systems. The TMT is a better substitute for the DT than the IHAT.

Antibodies↗

Toxoplasmosis.

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Female↗

Age-sex distribution of various diseases with particular reference to toxoplasmic lymphadenopathy.

An account is given of some human diseases which affect one sex more than the other. An age-sex realtionship has been noted among British patients with acquired toxoplasmic lymphadenopathy. This is compared with the findings of other European workers. A possible explanation is offered taking all these diseases into consideration together with some of the experimental work done in animals and some of the variations in immunological responses by man.

Adolescent↗

A microplate enzyme-immunoassay for toxoplasma antibody.

A new test for the detection and measurement of toxoplasma antibody is described. Test sera are reacted with antigen-sensitized wells in micro-haemagglutination plates. Any attached antibody is shown by the addition of an enzyme-labelled antiglobulin followed by assay of the enzyme reaction with its substrate. The test is easy to carry out on a large scale, and there is a positive correlation between the results and dye test and haemagglutination test titres.

Alkaline Phosphatase↗