Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1968 Nov;96(5):1500–1506. doi: 10.1128/jb.96.5.1500-1506.1968

Reduction of Nonspecific Background Staining in the Fluorescent Treponemal Antibody-Absorption Test

Merritt E Roberts a, James N Miller a,1, Gerald F Binnings a
PMCID: PMC315201  PMID: 4177869

Abstract

The nonspecific background fluorescence which occurs with the fluorescent treponemal antibody-absorption test for syphilis was found to result from a reaction between serum-treated Treponema pallidum organisms and the conjugated antihuman γ-globulin. It was also shown that β-lipoprotein and albumin were the important contributing factors in human serum. Various dilutions of 2.5% trypsin in phosphate-buffered saline specifically reduced background fluorescence under proper test conditions. By employing a trypsin digestion method, a semiautomated procedure utilizing a visual readout has been postulated as feasible.

Full text

PDF
1505

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beutner E. H. IMMUNOFLUORESCENT STAINING: THE FLUORESCENT ANTIBODY METHOD. Bacteriol Rev. 1961 Mar;25(1):49–76. doi: 10.1128/br.25.1.49-76.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. DEACON W. E., FALCONE V. H., HARRIS A. A fluorescent test for treponemal antibodies. Proc Soc Exp Biol Med. 1957 Nov;96(2):477–480. doi: 10.3181/00379727-96-23512. [DOI] [PubMed] [Google Scholar]
  3. DEACON W. E., HUNTER E. F. Treponemal antigens as related to identification and syphilis serology. Proc Soc Exp Biol Med. 1962 Jun;110:352–356. doi: 10.3181/00379727-110-27515. [DOI] [PubMed] [Google Scholar]
  4. FIFE E. H., Jr, BRYAN B. M., SANDERS R. W., MUSCHEL L. H. Evaluation of the fluorescent treponemal antibody (FTA) test for syphilis. Comparison with Treponema pallidum immobilization (TPI) test. Am J Clin Pathol. 1961 Aug;36:105–113. doi: 10.1093/ajcp/36.2.105. [DOI] [PubMed] [Google Scholar]
  5. FOGEL B. J., SANDERS R. W., FIFE E. H., Jr, ANDERSON R. I. THE SERODIAGNOSIS OF EARLY SYPHILIS, HOW TO UTILIZE THE NEWER LABORATORY TESTS. Clin Pediatr (Phila) 1965 Aug;4:447–452. doi: 10.1177/000992286500400805. [DOI] [PubMed] [Google Scholar]
  6. Frommhagen L. H. The solubility and other physicochemical properties of human gamma-globulin labeled with fluorescein isothiocyanate. J Immunol. 1965 Sep;95(3):442–445. [PubMed] [Google Scholar]
  7. GEORGALA D. L., BOOTHROYD M. PREPARATION OF FLUORESCENT POLYVALENT SALMONELLA ANTISERA. Nature. 1965 Jan 30;205:521–522. doi: 10.1038/205521a0. [DOI] [PubMed] [Google Scholar]
  8. HUNTER E. F., DEACON W. E., MEYER P. E. AN IMPROVED FTA TEST FOR SYPHILIS, THE ABSORPTION PROCEDURE (FTA-ABS). Public Health Rep. 1964 May;79:410–412. [PMC free article] [PubMed] [Google Scholar]
  9. Hebert G. A., Pittman B., Cherry W. B. Factors affecting the degree of nonspecific staining given by fluorescein isothiocyanate labelled globulins. J Immunol. 1967 Jun;98(6):1204–1212. [PubMed] [Google Scholar]
  10. LABREC E. H., FORMAL S. B., SCHNEIDER H. Serological identification of Shigella flexneri by means of fluorescent antibody. J Bacteriol. 1959 Sep;78:384–391. doi: 10.1128/jb.78.3.384-391.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LEIBOVITZ A., OBERHOFER T. R., MEACHAM J. T., Jr, DIESTELHORST T. N. ENHANCEMENT OF SPECIFICITY OF THE FLUORESCENT TREPONEMAL ANTIBODY TEST AS COMPARED WITH THE TPI TEST. Am J Clin Pathol. 1963 Nov;40:480–486. doi: 10.1093/ajcp/40.5.480. [DOI] [PubMed] [Google Scholar]
  12. LEVINE S. I., GOULET N. R., LIU O. C. BETA-PROPIOLACTONE DECONTAMINATION OF SIMIAN VIRUS-40 AS DETERMINED BY A RAPID FLUORESCENT-ANTIBODY ASSAY. Appl Microbiol. 1965 Jan;13:70–72. doi: 10.1128/am.13.1.70-72.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LEWIS V. J., JONES W. L., BROOKS J. B., CHERRY W. B. TECHNICAL CONSIDERATIONS IN THE PREPARATION OF FLUORESCENT-ANTIBODY CONJUGATES. Appl Microbiol. 1964 Jul;12:343–348. doi: 10.1128/am.12.4.343-348.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Levinthal J. D., Ahmad-Zadeh C., Van Hoosier G., Jr, Trentin J. J. Immunofluorescence of human adenovirus type 12 in various cell types. Proc Soc Exp Biol Med. 1966 Feb;121(2):405–414. doi: 10.3181/00379727-121-30791. [DOI] [PubMed] [Google Scholar]
  15. MARSHALL J. D., EVELAND W. C., SMITH C. W. Superiority of fluorescein isothiocyanate (Riggs) for fluorescent-antibody technic with a modification of its application. Proc Soc Exp Biol Med. 1958 Aug-Sep;98(4):898–900. doi: 10.3181/00379727-98-24222. [DOI] [PubMed] [Google Scholar]
  16. Pittman B., Herbert G. A., Cherry W. B., Taylor G. C. The quantitation of nonspecific staining as a guide for improvement of fluorescent antibody conjugates. J Immunol. 1967 Jun;98(6):1196–1203. [PubMed] [Google Scholar]
  17. Stout G. W., Kellogg D. S., Jr, Falcone V. H., McGrew B. E., Lewis J. S. Preparation and standardization of the sorbent used in the fluorescent treponemal antibody-absorption (FTA-ABS) test. Health Lab Sci. 1967 Jan;4(1):5–8. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES