Treponema Pallidum Immobilization Research Articles

The antigenic interrelationship between the endoflagella of Treponema phagedenis biotype Reiter and Treponema pallidum Nichols strain. I. Treponemicidal activity of cross-reactive endoflagellar antibodies against T. pallidum.

Treponemicidal activity against Treponema pallidum, Nichols strain, by anti-endoflagellar antibodies and the presence of antigenic interrelationships between the endoflagella of Treponema phagedenis biotype Reiter (TPR) and T. pallidum have been demonstrated. SDS-PAGE profiles of purified endoflagella from both organisms were similar, identifying five polypeptide bands for TPR (37,000, 33,000 doublet, 30,000, and 27,000 daltons) and five polypeptide bands for T. pallidum (35,000, 33,000 doublet, 30,000, and 27,000 daltons). Antiserum against TPR endoflagella identified identical bands on Western blots of TPR, T. pallidum, and the respective endoflagellar preparations. Western blots confirmed the presence of antibodies in normal human serum (NHS) against the 33,000 dalton treponemal endoflagellar proteins. The complement-dependent treponemicidal activity of NHS against T. pallidum was completely removed by absorption with purified TPR endoflagella. Furthermore, rabbit antisera against TPR endoflagella were reactive in the Treponema pallidum immobilization (TPI) test. These findings demonstrate that anti-endoflagellar antibodies are treponemicidal against T. pallidum. A possible mechanism for this activity is discussed in relation to the subsurface location of endoflagella.

Failure of the Treponema pallidum immobilization test to provide additional diagnostic information about contemporary problem sera.

Two hundred forty-five sera submitted to the Center for Disease Control, Atlanta, Ga., (CDC) were analyzed serologically in an attempt to demonstrate the diagnostic value of the Treponema pallidum immobilization (TPI) test when performed in addition to the fluorescent treponemal antibody-absorption (FTA-Abs) test. Diagnoses for the patients whose sera were tested were based on information supplied by the referring physicians. Fifty-four per cent of the diagnostic problems were resolved merely by the finding of a negative result with the FTA-Abs test. There was agreement between the serologic results of the referring laboratory and those of the CDC for 76% of sera tested by the Venereal Disease Research Laboratory test and for 71% of sera tested by the FTA-Abs test. For patients with treponemal disease, the sensitivity of the TPI test was 56% and that of the FTA-Abs test was 78%. When the FTA-Abs test was reactive, a negative TPI test was not significantly associated with systemic lupus erythematosus (P > 0.6) or other collagen vascular disease (P > 0.6), nor was a positive TPI test associated with treponemal disease (P > 0.09). It is concluded that once the result of the FTA-Abs test is known, the TPI test does not provide additional diagnostic information.

Cerebrospinal fluid findings in asymptomatic patients with reactive serum fluorescent treponemal antibody absorption tests.

It is common to examine the cerebrospinal fluid in untreated or inadequately treated asymptomatic patients with a reactive serum fluorescent treponemal antibody absorption (FTA-ABS) test before initiating antibiotic therapy for syphilis. This prospective study evaluated the usefulness of such examination. Four hundred thirty-two patients over 40 years old, reporting for annual physical examination, had a serum FTA-ABS test. Thirty-seven (8.6%) patients and 2 of 4 spouses were reactive repeatedly. Of the 39 patients with reactive tests, 7 had a history of penicillin therapy for syphilis, 5 had received heavy metal therapy, and 27 had no history of syphilis. These 39 patients had a neurological examination, serum VDRL, Treponema pallidum immobilization (TPI), and repeat FTA-ABS tests by two other laboratories. The TPI test was reactive in 30 (77%). Four had nonspecific neurological signs. Routine CSF examination (cells, total protein, VDRL, glucose, IgG%) on 30 patients with a history of inadequate treatment had a low diagnostic yield. Two patients had an unexplained total protein elevation (57 and 61 mg/dl) and 1 had a mildly increased IgG% (15%). All cell counts, VDRL tests, and glucose levels were normal. Agarose electrophoresis demonstrated one or more CSF immunoglobulin bands in 10 (36%) of 28 patients, possibly representing an immunological marker of past or latent central nervous system infection.

Borderline and reactive FTA-ABS. Results in lupus erythematosus

Fluorescent treponemal antibody absorption tests were performed on serum samples from 43 patients with lupus erythematosus (LE). Seven borderline and three reactive results were obtained from patients who had no evidence of syphilis, and Treponema pallidum immobilization (TPI) tests were nonreactive. These probable nonsyphilitic reactions were associated with discoid LE (DLE) as frequently as with systemic LE (SLE). Three of these serum samples were reactive in the microhemagglutination assay for T pallidum antibodies (MHA-TP), but titers were low. Reactive FTA-ABS tests were also obtained from four patients with LE with evidence of syphilis. All four serum samples were highly reactive in the MHA-TP test. These findings suggest that nonsyphilitic FTA-ABS fluorescence is (1) common with both DLE and SLE, (2) frequently not "beaded" in pattern, and (3) often detectable when MHA-TP tests are performed.

Late syphilis. Otologic symptoms and results of the FTA-ABS test.

The clinician occasionally encounters a patient whose cochleovestibular symptoms are thought to be due to late syphilis mainly on the basis of a reactive FTA-ABS test. I describe 21 such patients, 11 of whom had either a spurious- or a false-positive reaction. The following suggestions may avoid an inaccurate diagnosis of late syphilis in such patients: (1) repeat all reactive serological tests for syphilis (STS); (2) rule out false-positive reactions; (3) request the degree of fluorescence (1+ to 4+) on all reactive FTA-ABS tests; (4) obtain a Treponema pallidum immobilization (TPI) test on all patients with repeatedly reactive (1+) FTA-ABS tests--a nonreactive TPI test rules out the diagnosis of syphilis; (5) if a TPI test cannot be performed, a repeatedly reactive (1+) FTA-ABS test in the absence of a reactive nontreponemal STS probably represents a false-positive reaction; (6) clinical judgment is the final determinant of whether a patient has or should be treated for late syphilis.

Immunity in Experimental Syphilis

Abstract Twenty-four rabbits, each immunized intravenously over a 37-week period with a total of 3.71 × 109 γ-irradiated, non-infectious Treponema pallidum, Nichols strain, were found to be completely resistant to intradermal challenge with either 1000 or 100,000 homologous treponemes administered 10 days after the last immunizing injection. Persistence of complete immunity was demonstrable in each of 11 vaccinated rabbits challenged intradermally with 1800 T. pallidum, Nichols strain, 1 year after the last immunizing injection. In contrast, only minimal resistance was observed in 10 of the T. pallidum-immunized rabbits challenged intradermally with 1000 Treponema pertenue, Haiti B strain, 14 days after completion of immunization. Venereal Disease Research Laboratory, Treponema pallidum immobilization (TPI), and Fluorescent Treponemal Antibody-Absorbed antibodies, each of which is important in the diagnosis and control of syphilis, developed during immunization and persisted in some animals for at least 1 year. Conclusive evidence that TPI antibody is not associated with the immune response was provided by the finding that 8 of the 11 immune rabbits challenged 1 year after vaccination had no TPI antibody before challenge and failed to develop immobilizing antibody during the 3-month period of observation after challenge.

Evaluation of the Qualitative and Automated Quantitative Microhemagglutination Assay for Antibodies to Treponema pallidum

Qualitative and quantitative microhemagglutination assays for antibodies to Treponema pallidum (MHA-TP) were performed on 314 syphilitic and 597 presumably nonsyphilitic sera, and the results were compared with those of the fluorescent treponemal antibody-absorbed (FTA-ABS), the Treponema pallidum immobilization (TPI), and the Veneral Disease Research Laboratory (VDRL) tests. MHA-TP sensitivity was similar to that of the other tests in all stages of syphilis except primary syphilis, in which MHA-TP reactivity was only 64% compared with 82% in the FTA-ABS test, 73% in the VDRL test, and 67% in the TPI test. MHA-TP specificity was satisfactory and comparable to that of the other treponemal tests. Quantitation of the MHA-TP test was automated by use of Autotiter II equipment. Titers tended to become elevated later in the course of syphilis and to remain elevated longer than did VDRL titers. Reproducibility of the quantitative MHA-TP test was satisfactory, with duplicate tests agreeing within one doubling dilution on 97.5% of 351 reactive sera. Poor reproducibility was obtained with sera giving minimal reactions in the qualitative test, and such sera should be routinely retested. The MHA-TP is less time-consuming and costly than the FTA-ABS test and could be used in conjunction with the VDRL or another reagin test for syphilis to eliminate a large number of the FTA-ABS tests now required.

Spirochetes in Late Seronegative Syphilis, Penicillin Notwithstanding.

Before the advent of serological tests for syphilis, the disease was difficult to diagnose. When Wasserman, Hinton, and others developed their serological tests, the diagnostic problem was simplified. With the development of Reiter protein complement fixation (RPCF), fluorescent treponemal antibody (FTA), Treponema pallidum immobilization (TPI), and fluorescent treponemal antibodyabsorption (FTA-ABS) tests, both the specificity and sensitivity have been greatly improved. What is most conclusive, however, is the demonstration of Treponema pallidum , not only during the early stages but also at the late stage of the disease. With the advent of penicillin, treatment of syphilis has become a simple matter. Nowadays, syphilis is considered as just another infectious disease, no longer taught separately as syphilology. Nonetheless, in this book, J. L. Smith takes the opposite view: syphilis is now both hard to diagnose and hard to treat. Section 1 of the book reviews the problem of late seronegative (by reaginic tests)

Early Syphilis: Immunoglobulins Reactive in Immunofluorescence and other Serologic Tests

Summary Sera and serum fractions from 41 patients with darkfield-positive untreated primary and secondary syphilis were studied to reveal the immunoglobulins that are reactive in immunogluorescence and other serologic tests for syphilis. A monospecific indirect fluorescent antibody (IFA) assay revealed that IgG is the major immunoglobulin class reactive with Treponema pallidum, and that smaller amounts of IgM and IgA antibodies may also react. IgG is the principal immunoglobulin operative in the fluorescent treponemal antibody-absorption (FTA-ABS) test, and in its predecessor tests, the fluorescent treponemal antibody 1:5 (FTA-1:5), and the fluorescent treponemal antibody-200 (FTA-200). “Sorbent,” or a sonicate of Reiter treponemes, which is used in the FTA-ABS test to distinguish syphilitic from nonsyphilitic sera, operationally acts mainly to prevent “natural” IgG antibodies in nonsyphilitic serum from reacting with T. pallidum. It does not prevent syphilitic IgG antibodies from reacting, and apparently it enhances their role relative to IgM and IgA antibodies. Reactivity in the Treponema pallidum immobilization (TPI) test and the Reiter protein complement-fixation (RPCF) test was found mainly in the 7 S fractions of sera as compared to the 19 S fractions. Absorbing a syphilitic serum with T. pallidum did not produce a measurable decrease in total levels of IgG, IgM, or IgA. Antilipoidal antibodies detectable in the VDRL slide test were found in both the 19 S and 7 S fractions of sera; the antibodies associated with primary syphilis were not confined to the IgM class. In the course of these studies, 19 S and 3.5 S IgG-related substances were noted to possess anti-T. pallidum reactivity, and IgA in the 7 S fraction of serum was observed to be functionally silent in contrast to IgA of higher weight.

Serodiagnosis of Syphilis

To the Editor:— We are in general agreement with the answer of your consultant regarding the handling of a 54-year-old housewife having a weekly reactive venereal disease research laboratory (VDRL) slide test and a reactive fluorescent treponemal antibody-absorption procedure with serum (FTA-ABS) test ( 203 : 996, 1968). Evidence available to me indicates that in the serodiagnosis of syphilis the FTA-ABS test is as specific as the Treponema pallidum immobilization (TPI) test, yet more sensitive. Of course, patients having yaws or pinta, or other treponematoses closley related to syphilis, may also be expected to react; but these diseases would not usually be considered in the United States unless the patient has lived or traveled in countries where the diseases are endemic. A subsequent correspondent ( 204 : 833, 1968) agreed with your consultant's emphasis on the possibility or probability of syphilis, but also urged that efforts be made to identify factors which might be

The TPI and FTA-ABS Tests in Treated Late Syphilis

Sixty-seven patients who had been diagnosed and treated 13 or more years ago for latent or late syphilis were retested with use of several serological tests for syphilis. At the time of original diagnosis, all patients had been reactive in cardiolipin reagin tests and in the Treponema pallidum immobilization (TPI) test. It was found that 73% of patients were still reactive in one or more cardiolipin reagin tests and that 90% had retained TPI reactivity. In the newer fluorescent treponemal antibody-absorption (FTA-ABS) test, 98% were reactive. These results indicate that the FTA-ABS test is more sensitive than the TPI test. Neither the FTA nor the TPI test, as presently performed, may be used as an index of therapeutic response, since both tests show reactivity 13 or more years after persumably adequate treatment .

The TPI and FTA-ABS tests in treated late syphilis.

Sixty-seven patients who had been diagnosed and treated 13 or more years ago for latent or late syphilis were retested with use of several serological tests for syphilis. At the time of original diagnosis, all patients had been reactive in cardiolipin reagin tests and in the Treponema pallidum immobilization (TPI) test. It was found that 73% of patients were still reactive in one or more cardiolipin reagin tests and that 90% had retained TPI reactivity. In the newer fluorescent treponemal antibody-absorption (FTA-ABS) test, 98% were reactive. These results indicate that the FTA-ABS test is more sensitive than the TPI test. Neither the FTA nor the TPI test, as presently performed, may be used as an index of therapeutic response, since both tests show reactivity 13 or more years after persumably adequate treatment .

THE VALUE OF THE TREPONEMA PALLIDUM IMMOBILIZATION TEST (T.P.I.) IN ANOMALOUS SYPHILIS SEROLOGY

Medical Journal of AustraliaVolume 2, Issue 23 p. 1101-1103 Reports of Case THE VALUE OF THE TREPONEMA PALLIDUM IMMOBILIZATION TEST (T.P.I.) IN ANOMALOUS SYPHILIS SEROLOGY M. F. Garner M.B., B.S., M.C.Path., M. F. Garner M.B., B.S., M.C.Path. The Institute of Clinical Pathology and Medical Research, Department of Public Health, Sydney1 Serologist.Search for more papers by this authorN. M. Grantham B.Sc., N. M. Grantham B.Sc. The Institute of Clinical Pathology and Medical Research, Department of Public Health, Sydney2 Microbiologist.Search for more papers by this author M. F. Garner M.B., B.S., M.C.Path., M. F. Garner M.B., B.S., M.C.Path. The Institute of Clinical Pathology and Medical Research, Department of Public Health, Sydney1 Serologist.Search for more papers by this authorN. M. Grantham B.Sc., N. M. Grantham B.Sc. The Institute of Clinical Pathology and Medical Research, Department of Public Health, Sydney2 Microbiologist.Search for more papers by this author First published: 01 December 1966 https://doi.org/10.5694/j.1326-5377.1966.tb91857.x 3 To conform to WHO recommendations, sera are described as “reactive” and “non-reactive”, instead of as “positive” and “negative”. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume2, Issue23December 1966Pages 1101-1103 RelatedInformation

Serologic Tests for Syphilis and the False-Positive Reactor

The role of treponemal and nontreponemal tests in the serodiagnosis of syphilis is reviewed. Diagnostic problem cases should be evaluated by treponemal tests to distinguish latent syphilis from false-positive reactions. A comparison of treponemal tests performed on serum from 101 late latent and tertiary syphilitics showed that the fluorescent treponemal antibody absorption (FTA-ABS) test is more sensitive than the Treponema pallidum immobilization (TPI) or FTA-200 tests. Objective evidence of late syphilis was present in 6 of 11 patients with discordant test results (TPI nonreactive; FTA-ABS reactive). Of the 50 chronic false-positive reactors studied, 24 had evidence of lupus erythematosus or another of the autoimmune diseases.

Laboratory studies in retinitis pigmentosa.

Serum cholesterol levels are said to be lower in the Bassen-Kornzweig syndrome (acanthocytosis, ataxia, lipid abnormalities, and retinitis pigmentosa) than in any other known clinical entity (less than 75 mg/100 cc). 1 This statement prompted the following study of blood lipids in adult patients with retinitis pigmentosa. Since syphilitic chorioretinitis can so closely imitate retinitis pigmentosa, 2 even in terms of an extinguished electroretinogram (ERG), 3 and because many cases of seronegative ocular syphilis can be serologically detected only by the treponema pallidum immobilization (TPI) and fluorescent treponemal antibody absorption (FTA-ABS) tests, 4 each patient in this study received a TPI and FTA-ABS test as well as an ERG. Because inconsistent laboratory values have been reported in cases of retinitis pigmentosa, serum vitamin A, carotene, protein electrophoresis, red cell morphology, and latex flocculation, as well as urinary amino acids, were also evaluated. This report compares data obtained from nine adult

Evaluation of fluorescent treponemal antibody (RFTA and FTA II) and other tests (RPCF and TPI) for syphilis.

SummaryFluorescent treponemal antibody tests (RFTA and FTA II) using hypochlorite-treated Reiter and Nichols strain treponemes, respectively, were compared with each other and with a Reiter protein complement-fixation (RPCF) test in a study of sera previously subjected to the Treponema pallidum immobilization (TPI) test. The fluorescent antibody (FA) tests were closely comparable in sensitivity and specificity for syphilis. The RPCF test was slightly more sensitive than either FA test, but yielded somewhat more “non-specific” reactions. It appeared that either FA test might serve as well as the RPCF test as a screening procedure in differentiating persons with latent syphilis from those whose sera react “non-specifically” with cardiolipin antigens. Considerations favoring selection of an FA test are simplicity of performance and the need for fewer reagents. Moreover, the RFTA test substitutes an organism that can be cultivated simply in vitro for one requiring maintenance in infected rabbits.

Control Serum for the TPI Test

T HE Treponema pallidum immobilization (TPI) test described by Nelson (1,2) has been modified by many of the laboratories which perform this test (3). Controls for the originally described TPI test included a quantitatively titered syphilitic serum. At a meeting called by the Division of Venereal Disease, Public Health Service, in Washington, D.C., February 1, 1952, to discuss the TPI test, several participants expressed the need for a common, reactive serum, available to all laboratories, by which sensitivity levels of the TPI test could be measured from day to day within a laboratory and also between laboratories. Later that year such a control serum was distributed by the Public Health Service's Venereal Disease Experimental Laboratory to the Venereal Disease Research Laboratory and to other laboratories. This serum was frozen and sent by airmail in packages with solid CO2. Experiments conducted in 1952 (4) indicated that whole serum, to be used as quantitative reactive controls for the TPI test, could be distributed to distant laboratories in the liquid state without added preservatives or refrigeration. In March 1953 freeze-dried ampoules of the first international TPI control serum were distributed to 25 laboratories by the World Health Organization (5), and in February 1954 ampoules of the second international control serum were sent to 35 laboratories. Results of testing with these serums are reported and analyzed in a 1956 WHO report (6) in which the following statement is made regarding the use of a quantitative serum in the TPI test: In accordance with experiences from studies on reagin tests, the use of a reference (or standard) serum is recommended. In this way the effect of the variations of the titer levels between the laboratories as well as the temporal time variations within laboratories may be reduced. At the Venereal Disease Research Laboratory a reactive serum has been used as a quantitative control for the TPI test since July 1952. Since then the serum has been available to all TPI testing laboratories in the United States, under the following conditions: * A single serunm is to be common to all TPI testing laboratories, and this serum is to be replaced later by another serum that will be sent to all participating laboratories at the same time. The supply of serum on hand will suffice for at least 2 years. * All participating laboratories, including the Venereal Disease Research Laboratory, will keep a record of quantitative results obtained with this control serum each time it is used. A report containin-g these findings will be forwarded to the Venereal Disease Research Laboratory twice each year, in July and in January, where it will be duplicated and forwarded to all other participating laboratories without analysis or comment. The Venereal Disease Research Laboratory mailed the first shipments of serum, in quantities sufficient for approximately 6 months, to eight laboratories on January 20, 1958. The distribution of the serum is a continuing function of the Venereal Disease Research Laboratory, but only the results of testing for the 21/2-year period January 1958-June 1960 are included in this report, since a single lot of Afr. Harris and Dr. Brown are with the Communicable Disease Center, Public Health Service,, Atlanta, Ga. Mr. Harris is director, Venereal Disease Research Laboratory, Venereal Disease Branch, and Dr. Brown is chief of the branch.

Long-term serologic studies with treponemal procedures.

The purpose of this paper is to document our experience with treponemal testing since 1955 in a series of 1,064 clinically evaluated patients, and to discuss the patterns of seroreactivity after therapy in the various stages of syphilitic infection. We have been fortunate in being able to serologically evaluate certain cases for as long as 25 to 30 years after treatment. The tests used in this study consist of specific treponemal procedures as well as a battery of standard serologic tests for syphilis. The treponemal procedures employed were the Treponema pallidum complement-fixation 1 (TPCF) test, the Reiter protein complement-fixation 2 (RPCF) test, and the Treponema pallidum immobilization (TPI) test. All of the serums studied were subjected to the TPCF and RPCF tests in our laboratory. A small number of serums were tested with the TPI test which was performed by Dr. James N. Miller of the University of California Medical

Biologic False-Positive Reactions for Syphilis Among Narcotic Addicts

PRIOR to 1949, the occurrence of a reactive serologic test for syphilis (STS) in the absence of a history of lesions of the disease had been considered to be of equivocal significance by many investigators. In that year, the specific Treponema pallidum immobilization (TPI) test was developed by Nelson and Mayer 1 thus making available an accurate procedure for the differentiation of biologic false-positive (BFP) reactions from those due to infection with T. pallidum . Subsequently, numerous diseases and conditions have been observed to show BFP reactions, including collagen diseases, 2 pregnancy, 3 Brucella infection, and many others. During the past few years, increasing attention has been focused on the problem of narcotic addiction. Concomitant with this interest, it was observed that sera from many patients addicted to narcotics showed reactive STS. This report presents the incidence of BFP reactions among such individuals as measured by the TPI test. Methods

Comparison of the fluorescent treponemal antibody test with other tests for syphilis on cerebrospinal fluids.

Tests for syphilis on cerebrospinal fluids were evaluated in the Washington, D.C., Serology Conference of 1941 (J. vener. Dis. Inform., 1942), but spinal fluids were omitted from the later Serology Evaluation and Research Assembly Study in 1956-57 (S.E.R.A., 1959). Since this earlier evaluation, several treponemal antigen tests have been described. The Treponema pallidum immobilization (TPI), fluorescent treponemal antibody (FTA), and Kolmer complement-fixation tests using Reiter protein antigen (KRP) are three tests designed to detect antibodies differing from those that caused reactivity in the non-treponemal antigen tests. The FTA test has been used for testing sera, but has not properly been evaluated as a spinal fluid test. This study was conducted to evaluate the FTA test in comparison with other treponemal tests for syphilis on spinal fluids. The VDRL spinal fluid test was included in this study to offer a comparison of the non-treponemal with the treponemal test results.