1887

Abstract

Summary

Two sets of oligonucleotide primers were used to amplify the genomic DNA of , the causative agent of scrub typhus (tsutsugamushi disease), by the polymerase chain reaction. Each set of primers amplified 538-bp and 109-bp products, representing part of a gene encoding a possible major 58-kDa immunogenic protein, from whole genomic DNA extracted from strains Karp, Kato, Gilliam, Kuroki and Kawasaki. No amplification was observed from , mouse and human genomic DNA. DNA amplification was observed from crude lysates of peripheral whole blood, tissue homogenates and paraffin-embedded skin biopsy sections obtained from patients with scrub typhus disease. Southern blot analysis demonstrated the specificity of the amplified DNA fragments following hybridisation with a DNA probe generated from strain Karp. By means of this procedure, a rapid and sensitive diagnosis of scrub typhus disease can be made during the acute stage of this infection.

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/content/journal/jmm/10.1099/00222615-37-5-357
1992-11-01
2024-04-24
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References

  1. Eisemann C. S., Osterman J. V. Antigens of scrub typhus rickettsiae: separation by polyacrylamide gel electrophoresis and identification by enzyme-linked immunosorbent assay. Infect Immun 1981; 32:525–533
    [Google Scholar]
  2. Tamura A., Ohashi N., Urakami H., Takahashi K., Oyanagi M. Analysis of polypeptide composition and antigenic components of Rickettsia tsutsugamushi by polyacrylamide gel electrophoresis and immunoblotting. Infect Immun 1985; 48:671–675
    [Google Scholar]
  3. Hanson B. Identification and partial characterization of Rickettsia tsutsugamushi major protein immunogens. Infect Immun 1985; 50:603–609
    [Google Scholar]
  4. Urakami H., Ohashi N., Tsuruhara T., Tamura A. Characterization of polypeptides in Rickettsia tsutsugamushi: effect of preparative conditions on migration of polypeptides in polyacrylamide gel electrophoresis. Infect Immun 1986; 51:948–952
    [Google Scholar]
  5. Oaks E. V., Stover C. K., Rice R. M. Molecular cloning and expression of Rickettsia tsutsugamushi genes for two major protein antigens in Escherichia coli. Infect Immun 1987; 55:1156–1162
    [Google Scholar]
  6. Ohashi N., Tamura A., Ohta M., Hayashi K. Purification and partial characterization of a type-specific antigen of Rickettsia tsutsugamushi. Infect Immun 1989; 57:1427–1431
    [Google Scholar]
  7. Stover C. K., Marana D. P., Dasch G. A., Oaks E. V. Molecular cloning and sequence analysis of the Sta58 major antigen gene of Rickettsia tsutsugamushi: sequence homology and antigenic comparison of Sta58 to the 60-kilodalton family of stress proteins. Infect Immun 1990; 58:1360–1368
    [Google Scholar]
  8. Saiki R. K., Scharf S., Faloona F. Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 1985; 230:1350–1354
    [Google Scholar]
  9. Ou C.-Y., Kwok S., Mitchell S. W. DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells. Science 1988; 239:295–297
    [Google Scholar]
  10. Duggan D. B., Ehrlich G. D., Davey F. P. HTLV-1 induced lymphoma mimicking Hodgkin’s disease: diagnosis by polymerase chain reaction amplification of specific HTLV-1 sequences in tumor DNA. Blood 1988; 71:1027–1032
    [Google Scholar]
  11. Kaneko S., Miller R. H., Feinstone S. M. Detection of serum hepatitis B virus DNA in patients with chronic hepatitis using the polymerase chain reaction assay. Proc Natl Acad Sci USA 1989; 86:312–316
    [Google Scholar]
  12. Sambrook J., Fritsch E. F., Maniatis T. Molecular cloning: a laboratory manual. 2nd edn Cold Spring Harbor, NY: Cold Spring Harbor Laboratory; 1989
    [Google Scholar]
  13. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 1977; 74:5463–5467
    [Google Scholar]
  14. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983; 132:6–13
    [Google Scholar]
  15. Oaks E. V., Rice R. M., Kelly D. J., Stover C. K. Antigenic and genetic relatedness of eight Rickettsia tsutsugamushi antigens. Infect Immun 1989; 57:3116–3122
    [Google Scholar]
  16. Bozeman F. M., Elisberg B. L. Serological diagnosis of scrub typhus by indirect immunofluorescence. Proc Soc Exp Biol Med 1963; 112:568–573
    [Google Scholar]
  17. Tamura A., Takahashi K., Tsuruhara T. Isolation of Rickettsia tsutsugamushi antigenically different from Kato, Karp, and Gilliam strains from patients. Microbiol Immunol 1984; 28:873–882
    [Google Scholar]
  18. Eisemann C. S., Osterman J. V. Identification of strain-specific and group-reactive antigenic determinants on the Karp, Gilliam and Kato strains of Rickettsia tsutsugamushi. Am J Trop Med Hyg 1985; 34:1173–1178
    [Google Scholar]
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