Skip to main content
Clinical and Diagnostic Laboratory Immunology logoLink to Clinical and Diagnostic Laboratory Immunology
. 1994 Jan;1(1):44–50. doi: 10.1128/cdli.1.1.44-50.1994

Detection of borreliacidal antibody by using acridine orange and flow cytometry.

L C Lim 1, Y F Liu 1, K Schell 1, S D Lovrich 1, S M Callister 1, R F Schell 1
PMCID: PMC368194  PMID: 7496920

Abstract

Borreliacidal antibody has been shown to be important for the serodiagnosis of Lyme disease and determination of immune status. Our results show that borreliacidal antibody can be rapidly and accurately detected by flow cytometry. Acridine orange was added to normal and immune sera containing Borrelia burgdorferi organisms in the presence and absence of complement prior to data acquisition by flow cytometry. The flow cytometric parameters of side scatter and detection of acridine orange fluorescence were used to determine events per minute (number of labeled spirochetes), percent shift in fluorescence (number of dead spirochetes), and mean channel fluorescence (intensity of fluorescence-labeled spirochetes) of acridine orange-labeled spirochetes. Borreliacidal antibody was detected as early as 4 h, with optimal detection 16 to 24 h after incubation of B. burgdorferi organisms with immune serum and complement. Our results also showed that complement was necessary for detection of borreliacidal antibody. Flow cytometry with acridine orange-labeled spirochetes provides a rapid means for detection of borreliacidal antibody.

Full text

PDF
44

Selected References

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

  1. Bakken L. L., Case K. L., Callister S. M., Bourdeau N. J., Schell R. F. Performance of 45 laboratories participating in a proficiency testing program for Lyme disease serology. JAMA. 1992 Aug 19;268(7):891–895. [PubMed] [Google Scholar]
  2. Barbour A. G., Burgdorfer W., Grunwaldt E., Steere A. C. Antibodies of patients with Lyme disease to components of the Ixodes dammini spirochete. J Clin Invest. 1983 Aug;72(2):504–515. doi: 10.1172/JCI110998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Callister S. M., Case K. L., Agger W. A., Schell R. F., Johnson R. C., Ellingson J. L. Effects of bovine serum albumin on the ability of Barbour-Stoenner-Kelly medium to detect Borrelia burgdorferi. J Clin Microbiol. 1990 Feb;28(2):363–365. doi: 10.1128/jcm.28.2.363-365.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Callister S. M., Schell R. F., Case K. L., Lovrich S. D., Day S. P. Characterization of the borreliacidal antibody response to Borrelia burgdorferi in humans: a serodiagnostic test. J Infect Dis. 1993 Jan;167(1):158–164. doi: 10.1093/infdis/167.1.158. [DOI] [PubMed] [Google Scholar]
  5. Callister S. M., Schell R. F., Lovrich S. D. Lyme disease assay which detects killed Borrelia burgdorferi. J Clin Microbiol. 1991 Sep;29(9):1773–1776. doi: 10.1128/jcm.29.9.1773-1776.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Coleman J. L., Benach J. L. Isolation of antigenic components from the Lyme disease spirochete: their role in early diagnosis. J Infect Dis. 1987 Apr;155(4):756–765. doi: 10.1093/infdis/155.4.756. [DOI] [PubMed] [Google Scholar]
  7. Craft J. E., Grodzicki R. L., Steere A. C. Antibody response in Lyme disease: evaluation of diagnostic tests. J Infect Dis. 1984 May;149(5):789–795. doi: 10.1093/infdis/149.5.789. [DOI] [PubMed] [Google Scholar]
  8. Dressler F., Whalen J. A., Reinhardt B. N., Steere A. C. Western blotting in the serodiagnosis of Lyme disease. J Infect Dis. 1993 Feb;167(2):392–400. doi: 10.1093/infdis/167.2.392. [DOI] [PubMed] [Google Scholar]
  9. Grodzicki R. L., Steere A. C. Comparison of immunoblotting and indirect enzyme-linked immunosorbent assay using different antigen preparations for diagnosing early Lyme disease. J Infect Dis. 1988 Apr;157(4):790–797. doi: 10.1093/infdis/157.4.790. [DOI] [PubMed] [Google Scholar]
  10. Hechemy K. E., Harris H. L., Wethers J. A., Stevens R. W., Stock B. R., Reilly A. A., Benach J. L. Fluoroimmunoassay studies with solubilized antigens from Borrelia burgdorferi. J Clin Microbiol. 1989 Aug;27(8):1854–1858. doi: 10.1128/jcm.27.8.1854-1858.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hedberg C. W., Osterholm M. T., MacDonald K. L., White K. E. An interlaboratory study of antibody to Borrelia burgdorferi. J Infect Dis. 1987 Jun;155(6):1325–1327. doi: 10.1093/infdis/155.6.1325. [DOI] [PubMed] [Google Scholar]
  12. Karlsson M., Möllegård I., Stiernstedt G., Wretlind B. Comparison of Western blot and enzyme-linked immunosorbent assay for diagnosis of Lyme borreliosis. Eur J Clin Microbiol Infect Dis. 1989 Oct;8(10):871–877. doi: 10.1007/BF01963773. [DOI] [PubMed] [Google Scholar]
  13. Lane R. S., Lennette E. T., Madigan J. E. Interlaboratory and intralaboratory comparisons of indirect immunofluorescence assays for serodiagnosis of Lyme disease. J Clin Microbiol. 1990 Aug;28(8):1774–1779. doi: 10.1128/jcm.28.8.1774-1779.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lee D. A., Hoidal J. R., Clawson C. C., Quie P. G., Peterson P. K. Phagocytosis by polymorphonuclear leukocytes of Staphylococcus aureus and Pseudomonas aeruginosa adherent to plastic, agar, or glass. J Immunol Methods. 1983 Sep 30;63(1):103–114. doi: 10.1016/0022-1759(83)90213-2. [DOI] [PubMed] [Google Scholar]
  15. Lovrich S. D., Callister S. M., Schmitz J. L., Alder J. D., Schell R. F. Borreliacidal activity of sera from hamsters infected with the Lyme disease spirochete. Infect Immun. 1991 Aug;59(8):2522–2528. doi: 10.1128/iai.59.8.2522-2528.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Luger S. W., Krauss E. Serologic tests for Lyme disease. Interlaboratory variability. Arch Intern Med. 1990 Apr;150(4):761–763. [PubMed] [Google Scholar]
  17. Magnarelli L. A., Meegan J. M., Anderson J. F., Chappell W. A. Comparison of an indirect fluorescent-antibody test with an enzyme-linked immunosorbent assay for serological studies of Lyme disease. J Clin Microbiol. 1984 Aug;20(2):181–184. doi: 10.1128/jcm.20.2.181-184.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pantazis C. G., Kniker W. T. Assessment of blood leukocyte microbial killing by using a new fluorochrome microassay. J Reticuloendothel Soc. 1979 Aug;26(2):155–170. [PubMed] [Google Scholar]
  19. Pennell D. R., Wand P. J., Schell R. F. Evaluation of a quantitative fluorescence immunoassay (FIAX) for detection of serum antibody to Borrelia burgdorferi. J Clin Microbiol. 1987 Nov;25(11):2218–2220. doi: 10.1128/jcm.25.11.2218-2220.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Peterson P. K., Clawson C. C., Lee D. A., Garlich D. J., Quie P. G., Johnson R. C. Human phagocyte interactions with the Lyme disease spirochete. Infect Immun. 1984 Nov;46(2):608–611. doi: 10.1128/iai.46.2.608-611.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Russell H., Sampson J. S., Schmid G. P., Wilkinson H. W., Plikaytis B. Enzyme-linked immunosorbent assay and indirect immunofluorescence assay for Lyme disease. J Infect Dis. 1984 Mar;149(3):465–470. doi: 10.1093/infdis/149.3.465. [DOI] [PubMed] [Google Scholar]
  22. Sachsenmeier K. F., Schell K., Morrissey L. W., Pennell D. R., West R. M., Callister S. M., Schell R. F. Detection of borreliacidal antibodies in hamsters by using flow cytometry. J Clin Microbiol. 1992 Jun;30(6):1457–1461. doi: 10.1128/jcm.30.6.1457-1461.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schmitz J. L., Schell R. F., Hejka A. G., England D. M. Passive immunization prevents induction of Lyme arthritis in LSH hamsters. Infect Immun. 1990 Jan;58(1):144–148. doi: 10.1128/iai.58.1.144-148.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Steere A. C., Grodzicki R. L., Kornblatt A. N., Craft J. E., Barbour A. G., Burgdorfer W., Schmid G. P., Johnson E., Malawista S. E. The spirochetal etiology of Lyme disease. N Engl J Med. 1983 Mar 31;308(13):733–740. doi: 10.1056/NEJM198303313081301. [DOI] [PubMed] [Google Scholar]
  25. Wilske B., Schierz G., Preac-Mursic V., Weber K., Pfister H. W., Einhäupl K. Serological diagnosis of erythema migrans disease and related disorders. Infection. 1984 Sep-Oct;12(5):331–337. doi: 10.1007/BF01651147. [DOI] [PubMed] [Google Scholar]

Articles from Clinical and Diagnostic Laboratory Immunology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES