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Electron microscopy of treponemes subjected to the Treponema pallidum immobilization (TPI) test. I. Comparison of immunoimmobilized cells and control cells.

The ultrastructure of cells of T. pallidum Nichols subjected to the TPI test was studied in negatively stained specimens. Cells incubated in basal medium to which was added either human syphilis serum or heated guineapig serum (GPS) showed a normal morphology. This was also the cause for cells incubated with basal medium to which was added either human syphilis serum and heated GPS or normal human serum and unheated GPS. By dark-field microscopy cells obtained from these different incubation mixtures were found to be motile. In contrast, cells incubated in basal medium to which was added human syphilis serum and unheated GPS were all immobilized, and in the electron microscope they presented a morphology strikingly different from that of normal cells. The immunoimmobilized cells were swollen and their surface was completely covered with a layer of fuzzy material. The nature of this material and its possible role in rendering the treponemes immobile is discussed.

Animals

Electron microscopy of treponemes subjected to the Treponema pallidum immobilization (TPI) test. II. Immunoelectron microscopy.

The experiments were carried out in order to investigate whether human IgG globulin is absorbed on to the surface of T. pallidum cells during incubation with human syphilis serum in the Treponema pallidum immobilization (TPI) test. Cells of T. pallidum Nichols subjected to the TPI test were further incubated with ferritin conjugated rabbit antihuman IgG globulin. Human IgG globulin could only be demonstrated on immunoimmobilized cells, i.e. cells incubated with human syphilis serum and unheated guinea pig serum (GPS). The surface of the swollen cells was completely covered by a fuzzy layer on to which the ferritin molecules appeared to be attached. In ultrathin sections of cells obtained from the same suspensions the surface of the outer cell membrane was outlined by ferritin molecules. In these cells a rather wide gap was observed between the outer membrane and the cytoplasmic membrane. The ribosomes seemed to have disappeared from the cytoplasm of the immunoimmobilized treponemes, but were present in motile cells obtained from control incubations.

Adsorption

[Electroimmunodiffusion reaction in the diagnosis of syphilis (author's transl)].

A group of 800 human sera, obtained from syphilitic and nonsyphilitic individuals were tested in the counterimmunoelectrophoresis (CIE) technique with Reiter protein antigen. The sera were assayed for comparison in the following treponemal and nontreponemal tests: standard serologic tests for syphilis with cardiolipin; complement fixation (Kolmer 1/5 vol) with Reiter protein antigen (RPCF); T. pallidium immobilization (TPI) test; haemoagglutination test for T. pallidum antibodies (TPHA). The sensitivity and specificity of CIE test, its simplicity, the low cost, the possibility of practical application as a routine test for syphilis serology were discussed.

Antibodies

Comparison of cardiolipin and treponemal tests in the serodiagnosis of yaws.

Results of the Veneral Disease Research Laboratory (VDRL), rapid plasma reagin (RPR), Treponema pallidum haemagglutination (TPHA), T. pallidum immobilisation (TPI), and fluorescent treponemal antibody absorption (FTA-ABS) tests on sera of 661 children from a region where yaws is hypoendemic are compared. For 107 (16.2%) out of 661 sera the FTA-ABS test was the only one showing reactivity; in these instances the test was weakly reactive (intensity of fluorescence scored as +) and the children had no history and no signs or symptoms of treponemal disease. A solitary, weakly reactive FTA-ABS test result seems to have no clinical significance in these cases. The FTA-ABS test can be used as a confirmatory test for yaws instead of the TPI test, if only the results of sera showing an intensity of fluorescence scored as ++ or more are considered to be positive. There appeared to be no significant differences in the results of the VDRL, RPR, and TPHA tests as screening tests for yaws when the TPI or FTA-ABS tests were used as reference tests.

Adolescent

[The passive hemagglutination test in the diagnosis of syphilis].

Studies were carried out on the comparative value of passive haemaglutination, cardiolipin and treponema tests in a total number of 4327 sera syphilitic patients in various stages of the disease and apparently normal subjects or free of syphilis. The results demonstrated the specificity and sensitivity of passive haemagglutination in all the stages of syphilis. In view of the qualities and simplicity of the test, the author recommends it as a screening test combined with a micromethod using the cardiolipin antigen (flocculation test).

Complement Fixation Tests

[Passive haemagglutination test in serodiagnosis of syphilis. Survey of a one-year trial in a venereal diseases prophylactic center (author's transl)].

We report here a one-year study of the reliability of the Treponema pallidum Haemagglutination Test (TPHA) in the serological diagnosis of syphilis. By comparing TPHA data with the diagnosis based on a set of grouped criteria, we were able to determine TPHA accuracy. All studies were conducted at the Antivenereal Prophylactic Center, in Nice, France, on 1,775 sera. The grouped data consisted of:--Serological Test for Syphilis (STS) using cardiolipin antigen,--medical and epidemiological records;--two specific reactions; namely the Fluorescent Antibody Test (FTA) and the Treponema Immobilisation Test (TIT). Reliability is excellent in 98.58 p. 100: the TPHA either proves syphilis or eliminates this diagnosis. The TPHA was particularly useful in investigating "problem sera" for which there existed conflicts in the grouped data. The applicability of TPHA is limited, however, in that it has low sensitivity during early primary syphilis, and it cannot be used to monitor antibodies during treatment. Nevertheless, TPHA, based on a standard specific antigen, is assured of a high place among the tools of serological diagnosis of syphilis because of its high accuracy and sensitivity, its objective and easily read results, and the simplicity of its technique.

False Negative Reactions

Hemagglutination tests for syphilis antibody.

In a study of serodiagnosis of syphilis, the authors compared the specificities and sensitivities of two hemagglutination tests, a sheep-erythrocyte test (MHA-TP) and a trukey-erythrocyte test (TPHA), with those of the Fluorescent Treponemal Antibody-Absorption (FTA-ABS) test. In tests of sera from 935 patients without syphilis, the MHA-TP, TPHA, and FTA-ABS tests were reactive for 0.96, 0, and 1.3% respectively. The false-positive results were usually transient and not associated with underlying illness. For the 68 patients with syphilis, the MHA-TP test was as sensitive as the FTA-ABS test in all stages except untreated primary disease. The TPHA test appeared to be undersensitive, and testing of follow-up sera from persons with latent syphilis showed unexplained conversion of false-negative TPHA results to reactive results. Reproducibilities of the two hemagglutination tests were comparable. The MHA-TP test is a valuable confirmatory test for syphilis. Further study is needed before the use of the TPHA test can be recommended.

Animals

[Practical significance of syphilis serodiagnosis].

Recent developments in syphilis serology are set down. The new specific tests for treponema pallidum are described. Reactivity in different stages of the disease including congenital and neurosyphilis is discussed. An outline is given of the possibilities of the IgM-FTA-ABS and 19 S IgM-FTA-ABS tests. Syphilis serology today requires only 3 tests: a screening procedure, where the specific AMHA-TP is the method of choice, FTA-ABS test should be used for confirmation, and the quantitative VDRL in order to follow up the effectiveness of treatment.

Complement Fixation Tests