1887

Abstract

Summary

Dot hybridisation with P- and biotin-labelled probes prepared from leptospiral DNA was performed to develop a sensitive and specific diagnostic method for early infection with leptospires. The smallest amounts of leptospiral DNA that could be detected with P- and biotin-labelled probes was 1.5 pg and 5 pg, respectively, corresponding to about 750 and 2500 leptospires. Dot hybridisation with a P-labelled probe detected leptospiral DNA in sera from all of 14 experimentally infected golden hamsters. The smallest amount of leptospiral DNA detected in these experiments corresponded to about 2500 leptospires. In the test conditions described in this study, the sensitivity of dot hybridisation with a biotin-labelled probe was lower. Little cross-hybridisation was observed with unrelated DNAs which indicates that dot hybridisation could be a useful diagnostic method.

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/content/journal/jmm/10.1099/00222615-22-1-23
1986-08-01
2024-04-23
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References

  1. Brendle J. J., Rogul M., Alexander A. D. 1974; Deoxyribonucleic acid hybridization among selected leptospiral serotypes. International Journal of Systematic Bacteriology 24:205–214
    [Google Scholar]
  2. Buckel P., Zehelein E. 1981; Expression of Pseudomonas fluorescens D-galactose dehydrogenase in Escherichia coli. Gene 16:149–159
    [Google Scholar]
  3. Ellinghausen H. C., McCullough W. G. 1965; Nutrition of Leptospira pomona and growth of 13 other serotypes: a serum free medium employing oleic albumin complex. American Journal of Veterinary Research 26:39–44
    [Google Scholar]
  4. Haapala D. K., Rogul M., Evans L. B., Alexander A. D. 1969; Deoxyribonucleic acid base composition and homology studies of Leptospira. Journal of Bacteriology 98:421–428
    [Google Scholar]
  5. Johnson R. C., Harris V. G. 1967; Differentiation of pathogenic and saprophytic leptospires. I. Growth at low temperatures. Journal of Bacteriology 94:27–31
    [Google Scholar]
  6. Kohne D. E., Speicherwald A. G., Brenner D. J. 1984; Nucleic acid probe specific for members of the genus Legionella. In Thornsberry C. (eds) Legionella Proceedings of the second international symposium American Society for Microbiology; Washington D. C.:107–108
    [Google Scholar]
  7. Langer P. R., Waldrop A. A., Ward D. C. 1981; Enzymatic synthesis of biotin-labeled polynucleotides: Novel nucleic acid affinity probes. Proceedings of the National Academy of Sciences of the USA 78:6633–6637
    [Google Scholar]
  8. Leary J. J., Brigati D. J., Ward D. C. 1983; Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: bio-blots. Proceedings of the National Academy of Sciences of the USA 80:4045–4049
    [Google Scholar]
  9. Marshall R. B., Wilton B. E., Robinson A. J. 1981; Identification of lepsospira serovars by restriction-endonuclease analysis. Journal of Medical Microbiology 14:163–166
    [Google Scholar]
  10. Meinkoth J., Wahl G. 1984; Hybridization of nucleic acids immobilized on solid supports. Analytical Biochemistry 138:267–284
    [Google Scholar]
  11. Rigby P. W. J., Dieckmann M., Rhodes C., Berg P. 1977; Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  12. Ter Schegget J., Voves J., van Strien A., van der Noordaa J. 1980; Free viral DNA in BK virus-induced hamster tumor cells. Journal of Virology 35:331–339
    [Google Scholar]
  13. Wetmur J. G. 1975; Acceleration of DNA renaturation rates. Biopolymers 14:2517–2524
    [Google Scholar]
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