Biomedical subjects
Publications and source records attributed to H L Borkhardt.
The diagnostic importance of a new approach for the detection of N. go. by 1,2-Propanediol: NAD+-Oxidoreductase (1,2-PDOR) counting was demonstrated. The advantages are established in supporting of the culture diagnosis of cervical and urethral swabs and in the shorting of the diagnostic way by isolation of N. go. colonies and biochemical differentiation.
179 sera from patients with various stages of syphilis were investigated in the axial filament enzyme linked immunosorbent assay (AF-ELISA), the Treponema pallidum haemagglutination assay (TPHA), the fluorescence treponemal antibody absorption (FTA-ABS) test and the cardiolipin micro precipitation test (CMT). For the control of specificity 30 problem sera from non Treponema pallidum infected patients with a high quantity of Treponema genus specific antibodies and 40 sera from people without syphilis were also investigated. The sensitivity of the AF-ELISA (98.3%) was comparable with the TPHA (99.4%) and the FTA-ABS-test (98.9%). It was a high unspecificity in the group of problem sera.
Analysis by SDS-PAGE of axial filaments of cultivable treponemes showed 3 major bands: a 33/34 kD doublet and a 37 kD polypeptide. In sera from patients with various stages of syphilis, from biological false positive reactors, and from negative controls the most consistent reaction was detected against the doublet, but only sera from patients with primary or secondary syphilis reacted with the 37 kD axial filament polypeptides. About 6 weeks after antibiotic treatment sera had lost their reactivity with 37 kD antigens in agreement with decreasing antibodies in VDRL-test. Because of their recognition early in infection 37 kD-proteins are postulated to use in serodiagnostic assays, but our data emphasize also the need for use further antigens in tests for diagnosis in later stages of syphilis.
Hybridomas that secrete monoclonal antibody to Toxoplasma gondii have been developed. Two groups of 10 female BALB/c mice each were immunized either over a shorter (71 d) or longer (117 d) period at first with Toxoplasma lysate antigen and afterwards with intact tachyzoites of the RH strain. Higher titres of antibody were obtained with the long-period immunization. The fusion experiments have shown that both schemes of immunization approximately result in the same number (16 and 14% respectively) of hybridoma cell lines producing antigen-specific monoclonal antibodies. Hybridoma cultures secreting antitoxoplasma monoclonal antibodies were screened parallel by indirect immunofluorescence antibody test (IFAT) and enzyme immunoassay (EIA). 16 of the hybridoma cell cultures produced positive results only in the IFAT, 112 reacted only in the EIA and 21 were positive in both tests. The monoclonal antibodies 5B10, 5G6 and 1B2, which are positive in the IFAT form a chemical compound with the major antigens on the surface of RH tachyzoites. The patterns of fluorescence produced by these monoclonal antibodies are in conformity with those produced by using polyclonal sera of Toxoplasma gondii infected hosts (mouse, rabbit, man).
500 women in late pregnancy (III. trimenon) have been examined for colonization of group B streptococci by means of vaginal swabs. Identification of bacteria was effected with modified Wallerströmtest, CAMP-test and coagglutination. In 19 pregnant women (3.8%) B-streptococci could be found. In most cases type I was predominant, followed by type II and III. One newborn child died with symptoms of early onset disease. In mother and infant the same serotype could be found (III/R). The titre of maternal antibody, performed in B-streptococci ELISA was too low. A carrier with B-streptococci had a stillbirth. There existed a congenital abnormality. Both in mother and infant the serotype Ib/c was present. The titre of maternal antibody in B-streptococci ELISA was lowered.
The indirect enzyme-linked immunosorbent assay (ELISA) for detection of immunoglobulin M (IgM) antibodies to Treponema pallidum in sera of syphilitic patients is complicated by false positive reactions due to the interference of IgM rheumatoid factor (IgM-RF) activity and the presence of treponemal IgG antibodies. Another source of error producing false negative results is the competition between treponemal IgG and IgM antibodies for the binding sites on the antigen. To avoid these complications in the indirect Treponema pallidum-specific IgM-ELISA, total IgG was immunoprecipitated from sera of syphilitic patients prior to the assay. The IgM-RF from non-precipitated sera reacted in an IgM-RF-ELISA and in the Treponema pallidum-IgM-specific ELISA with identical titers. After precipitation of total IgG no reaction of the IgM-RF in the assay could be demonstrated. Competition between IgG and IgM antibodies can be prevented almost completely by the precipitation procedure. The sensitivity and specificity of the Treponema pallidum-specific IgM-ELISA after immunoprecipitation of total serum IgG were shown to be higher than 97 percent.
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In case of progressive paralysis of the insane, repeated recurrence can be explained only by the survival of pathogens inspite of optimal therapeutic conditions, the persistence of the pathogens being possibly produced by turning of the treponemas into L-type forms. In order that the possibilities and limits of recurrence verification will become apparent, reference is made to humoral and immunological changes of state given by way of example.
The examination tests annually necessary for a serological examination for syphilis and toxoplasmosis are in the first place as to their quantity in the diagnostics of the infection. For the performance of this large number of tests the use of a method is necessary which corresponds to the necessities of rationalisation of the laboratory work in the same way as to the demands to quality, which is characterized by specificity, sensitivity and reproducibility. For these criteria in the framework of the syphilis serology the TPHA-test is in a higher degree sufficient (with the exception of the early syphilis) than the FTA-ABS-test. The small quantity of the imported test set which is at present at our disposal does not allow a general application of this test, so that the FTA-ABS-test is the most usual method. Moreover, by means of FTA-ABS-IgM and 19 S-IgM-FTA-ABS-test valuable diagnostic additional informations (early diagnosis of the infection, L. connata diagnostics, estimation of the success of therapy) can be got which neither the TPI nor the TPHA-test are able to give. For the diagnosis of toxoplasmosis the IFT shows at present a higher value of precision than ELISA. It is still to be clarified in how far in the higher sensitivity of toxoplasmosis ELISA unspecificities play a role.
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