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E F Hunter

Publications and source records attributed to E F Hunter.

32 records · Page 2Linked to original sources
PubMed1976

Immunoglobulin specificity for the fluorescent treponemal antibody-absorption test conjugate.

Quantitative indirect immunofluorescence tests have demonstrated the predominance of immunoglobulin G antibody in most stages of syphilis. A class-specific anti-human immunoglobulin G conjugate is recommended to replace the present fluorescent treponemal antibody-absorption test conjugate. This would allow further definition and standardization of the fluorescent treponemal antibody-absorption test.

Antibody Specificity↗
PubMed1975

The specificity of fetal IgM: antibody or anti-antibody?

Reference materials were produced to standardize the immunoglobulin class specificity and potency of immunofluorescent anti-IgM conjugates used for diagnostic tests for congenital syphilis. In attempting to mimic essential immunologic characteristics of syphilitic and nonsyphilitic infant sera, we evaluated these sera in comparison with processed adult sera. We were quite surprised to discover that some syphilitic babies do not produce significant quantities to IgM antibody to T. pallidum in response to their infection, as would be expected; instead, they make relatively large amounts of IgM anti-IgG. We found this to be true also for newborns and infants infected with cytomegalovirus, rubella, and toxoplasmosis. To our knowledge, this observation has not been previously reported. However, it could have been predicted from the knowledge that older infants and young children normally produce IgM antibodies to maternal IgG allotypes (Gm factors). We are disturbed that these findings suggest that currently recommended indirect immunofluorescence IgM tests for perinatal infection may not be disease specific. Our observations may be important for a better understanding of basic immunologic mechanisms of fetal-maternal to tolerance and fetal response to life-threatening infection.

Adsorption↗
PubMed1975

The fluorescent treponemal antibody-absorption (FTA-ABS) test for syphilis.

The FTA test was developed at a time when immunofluorescence procedures were not well-defined. Through technique control and research, a modification of the FTA test, the FTA-ABS, has attained a position as one of the leading treponemal tests to confirm the reagin tests for syphilis. In this review of the FTA-ABS test, attention has been focused on reagent development, with the anticipation that reagent standardization may soon become a reality. The T. pallidum antigen obtained by extracting infected rabbit testicular tissue has evolved from a preparation in which the treponemes remained in the initial extracting fluid to a reagent that can be free of rabbit tissue and globulin. These washed antigen preparations improve visibility of the treponemes on the microscope slide, reduce background fluorescence, and reduce or prevent from occurring nonspecific reactions that are a result of tissue and globulin components. Both washed and nonwashed antigens are available commercially, and, to date, little differentiation has appeared on the product label. The predominant immunoglobulin that reacts with T. pallidum in the indirect fluorescent antibody tests appears to be IgG. This is the major immunoglobulin detected in the FTA-ABS test. IgM, although increased in early syphilis, is also increased in other clinical conditions. Several reports suggest that adult IgM detection in the present FTA-ABS test would be nonspecific. Until specific IgM antibody in adult syphilis can be detected without a risk to test specificity, the conjugate for the FTA-ABS test should continue to be an anti-IgG reagent. Class-specific, anti-IgG reagents are more expensive than other reagents; however, their use may eliminate the problem of nonspecificity resulting from IgM detection. Additionally, micromethods can be used to reduce cost, and this possibility should be investigated. The sorbent that contains an antigen to the Reiter treponeme may or may not specifically absorb the reactivity that occurs in normal sera; certainly, there are questionable aspects about this reagent. Group antibodies not related to Reiter treponemes may be responsible for some nonspecific reactivity; additionally, antiglobulin factors have been reported to participate in the reaction. Antigens free of rabbit serum factors and class-specific, antiimmunoglobulin reagents are available, and may lead to a better understanding of nonspecific reactions. These reagents should allow resolution of the possible multiplicity of reactivity. In this interim period, the sorbent, with its possible nonspecific nature, appears to maintain a biological balance between natural or group and immune antibodies when used to detect IgG antibody.

Absorption↗
PubMed1972

Immunochemical parameters of some commercial conjugates for the fluorescent treponemal antibody-absorption test.

Fluorescein-labeled anti-human globulins were examined to determine the need for standardization of conjugates used in the fluorescent treponemal antibody-absorption (FTA-ABS) test. Twenty-one of 33 conjugates submitted by commercial manufacturers to the Reagents Control Activity, Venereal Disease Research Laboratory, for evaluation in the FTA-ABS test were available for study. Conjugates, after evaluation in FTA-ABS performance tests, were examined by immunoelectrophoresis, by titration against immunoglobulins G and M (IgG, IgM) with FTA-ABS techniques, and by the biuret protein and fluorescein diacetate methods for determining fluorescein to protein (F/P) ratios. The conjugates were predominately anti-IgG globulin with anti-light-chain activity. Differences were noted in the ability of some conjugates to detect IgM antibody. The F/P ratios of those conjugates that could be determined varied from 2.6 to 17.8 mug of fluorescein per mg of protein. The need to identify and standardize both the immunologic capabilities and the optimum F/P ratio for FTA-ABS test conjugates is presented.

Animals↗
PubMed1968

The fluorescent treponemal antibody-absorption (FTA-ABS) test. Development, use and present status.

For testing problem sera, the fluorescent treponemal antibody-absorption test (FTA-ABS) is increasingly replacing other treponemal tests for syphilis (including the TPI test) in the USA. The authors review its development as a result of experience with other immunofluorescent techniques and discuss its advantages and disadvantages. Although it is a more sensitive indicator of past or present syphilitic infection than many other widely evaluated techniques, several aspects of the test are still not fully understood; in particular its specificity is not yet established and more information is needed on its reactivity in a variety of clinical diseases and in treponemal diseases other than syphilis. It is a relatively expensive technique in terms of technicians' time, reagents and equipment but there are grounds for hope that research now under way into automation of the procedure may improve the situation.

Absorption↗
PubMed1967

Antigenic relationships of 14 treponemes demonstrated by immunofluorescence.

Immunofluorescence was used as an aid in the antigenic grouping of 14 cultivable treponemes. Antisera were prepared versus each treponemal strain, and the antiglobulins were conjugated with fluorescein isothiocyanate. A common antigen-antibody system, detected in the strains studied, was removed by absorption of each conjugate with Reiter or Borrelia vincentii treponemes. Thus, five categories based on shared group-specific antigens were revealed. Serogroup I: Reiter, English Reiter, Kazan, Kazan numbers 2, 4, 5, and 8. Serogroup II: Nichols and Noguchi. Serogroup III: three oral treponemes, MRB, FM, and N-39. Serogroup IV: B. vincentii. Serogroup V: Treponema zuelzerae. The five serogroups apparently are related by an immunofluorescent common antigen.

Antigens↗
PubMedDate not supplied

New technologies for use in the surveillance and control of yaws.

In the Republic of Ghana, treponemal antigen tests performed on finger-prick blood from patients with yaws proved to be as sensitive as those tests performed on whole sera, and this mode of collection was more economical and acceptable than venipuncture. Under field conditions, dark-field microscopic examination of suspect yaws lesions was difficult as compared with collection of serous exudate in heparinized capillary tubes examined later in a reference laboratory. Direct staining of lesion exudate fixed on microscope slides with fluorescein-conjugated human or mouse monoclonal antibody against Treponema pallidum was more sensitive than dark-field examination. However, these techniques could not distinguish between the early lesions of venereal syphilis and those of yaws. An equally sensitive technique used a cloned segment of the T. pallidum (Nichols strain) genome to detect homologous DNA in lesion exudate fixed on nitrocellulose filter paper. The fixation of lesion exudates on microscope slides or nitrocellulose papers may prove to be the easiest method of collecting and transporting such materials to reference laboratories.

Adolescent↗
PubMedDate not supplied

Four serologic tests for syphilis: results with comparison of selected groups of sera.

The specificity of the fluorescent treponemal antibody-absorbed (FTA-Abs) test was assessed for 17 sera from syphilitic patients that were nonreactive in the Treponema pallidum immobilization (TPI) test but reactive in the FTA-Abs test. Thirty-three other sera from syphilitic patients and 19 sera from nonsyphilitic individuals were also examined by fluorescent treponemal and microhemagglutination Treponema pallidum (MHA-TP) tests and by the enzyme-linked immunosorbent assay (ELISA). Specific absorptions of sera with calf thymus DNA or Treponema pallidum biotype Reiter (Reiter treponemes) were performed. In quantitative immunofluorescence assays (IFA) with antihuman IgG and IgM conjugates, results were similar to those for reactive sera from a control group. Results of both the MHA-TP and ELISA tests supported the specificity of the FTA-Abs test; reactivity in the latter was not removed by specific absorption either with calf thymus DNA or with Reiter treponemes. This evaluation suggests a format for serodiagnosis in cases in which test results are discrepant.

Enzyme-Linked Immunosorbent Assay↗
PubMedDate not supplied

Evaluation of sera from patients with Lyme disease in the fluorescent treponemal antibody-absorption test for syphilis.

To determine whether the cross-reactivity between Treponema pallidum and Borrelia burgdorferi affects the specificity of the fluorescent treponemal antibody-absorption (FTA-Abs) test for syphilis, sera from patients with Lyme disease or syphilis were examined in a quantitative FTA-Abs test. Sera were diluted serially in phosphate-buffered saline, then in sorbent, and were tested with T. pallidum and B. burgdorferi antigens. Nine of 40 sera from patients with known Lyme disease were reactive at the 1:5 dilution with antigen from T. pallidum; only one serum was reactive at the 1:10 dilution. When both antigens were tested, the titer against B. burgdorferi was always higher than that against T. pallidum. Similarly, sera from patients with syphilis showed cross-reactivity with B. burgdorferi. Although reactivity could be absorbed with Treponemal phagedenis (Reiter strain), simultaneous titration with both antigens was easily performed and designated the etiologic agent.

Antigens, Bacterial↗
PubMedDate not supplied

Tissue fluid penetration and anti-treponemal activity of trospectomycin (a new spectinomycin analog) in the rabbit syphilis model.

The antitreponemal activity of trospectomycin (a novel 6'-propyl analog of spectinomycin) was correlated with concentrations in serum and tissue chamber fluid in a cutaneously-infected rabbit model of syphilis. After single-dose intramuscular (im) injections of trospectomycin, spectinomycin hydrochloride, and aqueous procaine penicillin G(APPG), concentrations of the drugs in paired specimens of serum and tissue fluid were determined and correlated with response of Treponema pallidum-infected lesions. Lesions responded most rapidly in APPG-treated animals. Rapid response correlated with more prolonged levels of APPG in both serum and tissue fluid. Trospectomycin, when given in single doses exceeding 40 mg/kg, surpassed penicillin in peak serum levels (greater than 60 micrograms/ml at 1 hr) but not in duration of activity. Higher and prolonged concentrations of trospectomycin in tissue fluid correlated with more effective clearance of treponemes from cutaneous lesions. Animals treated with less than 40 mg of trospectomycin/kg or with 20-80 mg/kg of the parent compound (spectinomycin hydrochloride) had cutaneous lesions that were persistently darkfield-positive, as confirmed by three or more consecutive smears.

Animals↗