Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1989 Sep;27(9):2073–2079. doi: 10.1128/jcm.27.9.2073-2079.1989

Line blot and western blot immunoassays for diagnosis of Mediterranean spotted fever.

D Raoult 1, G A Dasch 1
PMCID: PMC267741  PMID: 2506223

Abstract

The line blot, a new immunoassay in which antigens are placed on nitrocellulose as narrow lines, was evaluated for its sensitivity and specificity relative to the microimmunofluorescence assay for the diagnosis of Mediterranean spotted fever (MSF). The line blot assay was only slightly less sensitive and less specific than the microimmunofluorescence assay for detection of immunoglobulin M (IgM) or IgG in 100 serum specimens from 42 patients with MSF. No line blot reactions were observed among 50 control serum specimens from febrile patients with other illnesses. The line blot assay was largely group reactive for spotted fever rickettsiae, but 26% of the positive serum specimens also cross-reacted by IgM with Rickettsia typhi. Western immunoblotting was used to characterize the antigenic components recognized by 19 MSF serum specimens. For both IgM and IgG, lipopolysaccharide was the cross-reactive group antigen, whereas the high-molecular-weight species-specific protein antigens (SPAs) were the only reactive proteins. Relative to the other nine rickettsiae, Rickettsia bellii was unique both in exhibiting no SPA reactions and in having a lipopolysaccharide with a predominantly high-molecular-weight distribution. Although most of the 19 MSF serum specimens examined by Western blotting exhibited preferential reactivity to SPAs of two strains of R. conorii and weaker reactions to the other rickettsiae, 2 serum specimens exhibited SPA reactions consistent with typhus infections. In comparison with other assays, the line blot and Western blot immunoassays have advantages which may permit an improvement in the general availability and commercialization of assays for the serodiagnosis of rickettsial infections.

Full text

PDF
2073

Images in this article

2075Image
on p.2075
2076Image
on p.2076

Selected References

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

  1. Anacker R. L., List R. H., Mann R. E., Wiedbrauk D. L. Antigenic heterogeneity in high- and low-virulence strains of Rickettsia rickettsii revealed by monoclonal antibodies. Infect Immun. 1986 Feb;51(2):653–660. doi: 10.1128/iai.51.2.653-660.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anacker R. L., Mann R. E., Gonzales C. Reactivity of monoclonal antibodies to Rickettsia rickettsii with spotted fever and typhus group rickettsiae. J Clin Microbiol. 1987 Jan;25(1):167–171. doi: 10.1128/jcm.25.1.167-171.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dasch G. A. Isolation of species-specific protein antigens of Rickettsia typhi and Rickettsia prowazekii for immunodiagnosis and immunoprophylaxis. J Clin Microbiol. 1981 Sep;14(3):333–341. doi: 10.1128/jcm.14.3.333-341.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Devriendt J., Staroukine M., Amson R., Crockaert F., Dratwa M., Karmali R., Thoua Y. Malignant Mediterranean spotted fever. Report of a case with multiple organ failure, hypocalcemia, and euthyroid sick syndrome. Arch Intern Med. 1985 Jul;145(7):1319–1321. doi: 10.1001/archinte.145.7.1319. [DOI] [PubMed] [Google Scholar]
  5. Edlinger E. Serological diagnosis of Mediterranean spotted fever. Ann Microbiol (Paris) 1979 Feb-Mar;130(2):203–211. [PubMed] [Google Scholar]
  6. Feng H. M., Walker D. H., Wang J. G. Analysis of T-cell-dependent and -independent antigens of Rickettsia conorii with monoclonal antibodies. Infect Immun. 1987 Jan;55(1):7–15. doi: 10.1128/iai.55.1.7-15.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. García Curiel A., Nájera Morrondo E. Estudio epidemiológico de las rickettsiosis en la provincia de Sevilla, basado en las reacciones serológicas de inmunofluorescencia indirecta. Rev Sanid Hig Publica (Madr) 1984 Jan-Feb;58(1-2):83–98. [PubMed] [Google Scholar]
  8. Gear J. H., Miller G. B., Martins H., Swanepoel R., Wolstenholme B., Coppin A. Tick-bite fever in South Africa. The occurrence of severe cases on the Witwatersrand. S Afr Med J. 1983 May 21;63(21):807–810. [PubMed] [Google Scholar]
  9. Gross E. M., Goldwasser R. A., Bearman J. E., Sarov I., Sarov B., Torok V., Naggan L. Rickettsial antibody prevalence in southern Israel: IgG antibodies to Coxiella burnetii, Rickettsia typhi, and spotted fever group rickettsiae among urban- and rural-dwelling and Bedouin women. Am J Trop Med Hyg. 1983 Nov;32(6):1387–1391. doi: 10.4269/ajtmh.1983.32.1387. [DOI] [PubMed] [Google Scholar]
  10. Hechemy K. E., Anacker R. L., Philip R. N., Kleeman K. T., MacCormack J. N., Sasowski S. J., Michaelson E. E. Detection of Rocky Mountain spotted fever antibodies by a latex agglutination test. J Clin Microbiol. 1980 Aug;12(2):144–150. doi: 10.1128/jcm.12.2.144-150.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hechemy K. E. Laboratory diagnosis of Rocky Mountain spotted fever. N Engl J Med. 1979 Apr 12;300(15):859–860. doi: 10.1056/NEJM197904123001513. [DOI] [PubMed] [Google Scholar]
  12. Hechemy K. E., Raoult D., Eisemann C., Han Y. S., Fox J. A. Detection of antibodies to Rickettsia conorii with a latex agglutination test in patients with Mediterranean spotted fever. J Infect Dis. 1986 Jan;153(1):132–135. doi: 10.1093/infdis/153.1.132. [DOI] [PubMed] [Google Scholar]
  13. Hechemy K. E., Rubin B. B. Latex-Rickettsia rickettsii test reactivity in seropositive patients. J Clin Microbiol. 1983 Mar;17(3):489–492. doi: 10.1128/jcm.17.3.489-492.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hechemy K. E., Stevens R. W., Sasowski S., Michaelson E. E., Casper E. A., Philip R. N. Discrepancies in Weil-Felix and microimmunofluorescence test results for Rocky Mountain spotted fever. J Clin Microbiol. 1979 Feb;9(2):292–293. doi: 10.1128/jcm.9.2.292-293.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kaplan J. E., Schonberger L. B. The sensitivity of various serologic tests in the diagnosis of Rocky Mountain spotted fever. Am J Trop Med Hyg. 1986 Jul;35(4):840–844. doi: 10.4269/ajtmh.1986.35.840. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Li H., Lenz B., Walker D. H. Protective monoclonal antibodies recognize heat-labile epitopes on surface proteins of spotted fever group rickettsiae. Infect Immun. 1988 Oct;56(10):2587–2593. doi: 10.1128/iai.56.10.2587-2593.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mansueto S., Tringali G., Di Leo R., Maniscalco M., Montenegro M. R., Walker D. H. Demonstration of spotted fever group rickettsiae in the tache noire of a healthy person in Sicily. Am J Trop Med Hyg. 1984 May;33(3):479–482. doi: 10.4269/ajtmh.1984.33.479. [DOI] [PubMed] [Google Scholar]
  19. Mansueto S., Tringali G., Walker D. H. Widespread, simultaneous increase in the incidence of spotted fever group rickettsioses. J Infect Dis. 1986 Sep;154(3):539–540. doi: 10.1093/infdis/154.3.539-a. [DOI] [PubMed] [Google Scholar]
  20. Mansueto S., Vitale G., Miceli M. D., Tringali G., Quartararo P., Picone D. M., Occhino C. A sero-epidemiological survey of asymptomatic cases of Boutonneuse fever in western Sicily. Trans R Soc Trop Med Hyg. 1984;78(1):16–18. doi: 10.1016/0035-9203(84)90163-9. [DOI] [PubMed] [Google Scholar]
  21. McDonald G. A., Anacker R. L., Garjian K. Cloned gene of Rickettsia rickettsii surface antigen: candidate vaccine for Rocky Mountain spotted fever. Science. 1987 Jan 2;235(4784):83–85. doi: 10.1126/science.3099387. [DOI] [PubMed] [Google Scholar]
  22. Newhouse V. F., Shepard C. C., Redus M. D., Tzianabos T., McDade J. E. A comparison of the complement fixation, indirect fluorescent antibody, and microagglutination tests for the serological diagnosis of rickettsial diseases. Am J Trop Med Hyg. 1979 Mar;28(2):387–395. doi: 10.4269/ajtmh.1979.28.387. [DOI] [PubMed] [Google Scholar]
  23. Ormsbee R., Peacock M., Philip R., Casper E., Plorde J., Gabre-Kidan T., Wright L. Antigenic relationships between the typhus and spotted fever groups of rickettsiae. Am J Epidemiol. 1978 Jul;108(1):53–59. [PubMed] [Google Scholar]
  24. Philip R. N., Casper E. A., MacCormack J. N., Sexton D., Thomas L. A., Anacker R. L., Burgdorfer W., Vick S. A comparison of serologic methods for diagnosis of Rocky Mountain spotted fever. Am J Epidemiol. 1977 Jan;105(1):56–67. doi: 10.1093/oxfordjournals.aje.a112356. [DOI] [PubMed] [Google Scholar]
  25. Philip R. N., Casper E. A., Ormsbee R. A., Peacock M. G., Burgdorfer W. Microimmunofluorescence test for the serological study of rocky mountain spotted fever and typhus. J Clin Microbiol. 1976 Jan;3(1):51–61. doi: 10.1128/jcm.3.1.51-61.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Raoult D., De Micco C., Chaudet H., Tamalet J. Serological diagnosis of Mediterranean spotted fever by the immunoperoxidase reaction. Eur J Clin Microbiol. 1985 Aug;4(4):441–442. doi: 10.1007/BF02148712. [DOI] [PubMed] [Google Scholar]
  27. Raoult D., Hechemy K. E., Chaudet H. Sérologie de la fièvre boutonneuse méditerranéenne. Cinétique des anticorps détectés par trois méthodes: l'immunofluorescence indirecte, l'hémagglutination indirecte, et l'agglutination latex. Pathol Biol (Paris) 1985 Oct;33(8):839–841. [PubMed] [Google Scholar]
  28. Raoult D., Nicolas D., De Micco P., Gallais H., Casanova P. Aspects épidémiologiques de la fièvre boutonneuse méditerranéenne en corse du sud. Bull Soc Pathol Exot Filiales. 1985;78(4):446–451. [PubMed] [Google Scholar]
  29. Raoult D., Rousseau S., Toga B., Chaudet H., Tamalet J., de Micco P. Indirect fluorescent antibody test in Mediterranean spotted fever. Detection and kinetics of antibodies. Acta Virol. 1985 Dec;29(6):516–516. [PubMed] [Google Scholar]
  30. Raoult D., Rousseau S., Toga B., Tamalet C., Gallais H., De Micco P., Casanova P. Diagnostic sérologique de la fièvre boutonneuse méditerranéenne. Pathol Biol (Paris) 1984 Sep;32(7):791–794. [PubMed] [Google Scholar]
  31. Raoult D., Toga B., Chaudet H., Chiche-Portiche C. Rickettsial antibody in southern France: antibodies to Rickettsia conorii and Coxiella burnetii among urban, suburban and semi-rural blood donors. Trans R Soc Trop Med Hyg. 1987;81(1):80–81. doi: 10.1016/0035-9203(87)90290-2. [DOI] [PubMed] [Google Scholar]
  32. Raoult D., Weiller P. J., Chagnon A., Chaudet H., Gallais H., Casanova P. Mediterranean spotted fever: clinical, laboratory and epidemiological features of 199 cases. Am J Trop Med Hyg. 1986 Jul;35(4):845–850. doi: 10.4269/ajtmh.1986.35.845. [DOI] [PubMed] [Google Scholar]
  33. Raoult D., Zuchelli P., Weiller P. J., Charrel C., San Marco J. L., Gallais H., Casanova P. Incidence, clinical observations and risk factors in the severe form of Mediterranean spotted fever among patients admitted to hospital in Marseilles 1983-1984. J Infect. 1986 Mar;12(2):111–116. doi: 10.1016/s0163-4453(86)93508-5. [DOI] [PubMed] [Google Scholar]
  34. Raoult D., de Micco C., Gallais H., Toga M. Laboratory diagnosis of Mediterranean spotted fever by immunofluorescent demonstration of Rickettsia conorii in cutaneous lesions. J Infect Dis. 1984 Jul;150(1):145–148. doi: 10.1093/infdis/150.1.145. [DOI] [PubMed] [Google Scholar]
  35. Sompolinsky D., Boldur I., Goldwasser R. A., Kahana H., Kazak R., Keysary A., Pik A. Serological cross-reactions between Rickettsia typhi, Proteus vulgaris OX19, and Legionella bozemanii in a series of febrile patients. Isr J Med Sci. 1986 Oct;22(10):745–752. [PubMed] [Google Scholar]
  36. Weiss E., Coolbaugh J. C., Williams J. C. Separation of viable Rickettsia typhi from yolk sac and L cell host components by renografin density gradient centrifugation. Appl Microbiol. 1975 Sep;30(3):456–463. doi: 10.1128/am.30.3.456-463.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Westfall H. N., Goldwasser R. A., Weiss E., Hussong D. Prevalence of antibodies to Legionella species in a series of patients in Israel. Isr J Med Sci. 1986 Feb;22(2):131–138. [PubMed] [Google Scholar]

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

RESOURCES