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Probe directed at a segment of Rickettsia rickettsii rRNA amplified with polymerase chain reaction.

Publication ,  Journal Article
Wilson, KH; Blitchington, R; Shah, P; McDonald, G; Gilmore, RD; Mallavia, LP
Published in: J Clin Microbiol
December 1989

In an effort to explore a sensitive taxon-specific detection system for bacteria, we sequenced the 16S rRNA from two strains of Rickettsia rickettsii, five other rickettsiae, and Coxiella burnetti to find a probe site unique to R. rickettsii. We then synthesized a 16-mer that hybridized only to the rRNA of R. rickettsii. Using a primer complementary to a sequence found only in rickettsial rRNA, we then generated a cDNA. We amplified the probe site in a 180-base segment of the cDNA by using the cDNA primer and a second primer also unique to rickettsiae in a polymerase chain reaction. The segments of rRNA from each of the rickettsiae were amplified 10(6)- to 10(7)-fold, and the R. rickettsii probe hybridized only to the amplified segment from R. rickettsii. The rRNAs from Staphylococcus aureus, C. burnetii, and Neisseria meningitidis were not amplified and did not hybridize with the probe. The approach detailed below may prove clinically useful in the direct detection of pathogens that are difficult to cultivate.

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Published In

J Clin Microbiol

DOI

ISSN

0095-1137

Publication Date

December 1989

Volume

27

Issue

12

Start / End Page

2692 / 2696

Location

United States

Related Subject Headings

  • Staphylococcus aureus
  • Rickettsia typhi
  • Rickettsia rickettsii
  • Rickettsia prowazekii
  • Rickettsia
  • RNA, Viral
  • RNA, Ribosomal, 16S
  • RNA, Ribosomal
  • Polymerase Chain Reaction
  • Oligonucleotide Probes
 

Citation

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Wilson, K. H., Blitchington, R., Shah, P., McDonald, G., Gilmore, R. D., & Mallavia, L. P. (1989). Probe directed at a segment of Rickettsia rickettsii rRNA amplified with polymerase chain reaction. J Clin Microbiol, 27(12), 2692–2696. https://doi.org/10.1128/jcm.27.12.2692-2696.1989
Wilson, K. H., R. Blitchington, P. Shah, G. McDonald, R. D. Gilmore, and L. P. Mallavia. “Probe directed at a segment of Rickettsia rickettsii rRNA amplified with polymerase chain reaction.J Clin Microbiol 27, no. 12 (December 1989): 2692–96. https://doi.org/10.1128/jcm.27.12.2692-2696.1989.
Wilson KH, Blitchington R, Shah P, McDonald G, Gilmore RD, Mallavia LP. Probe directed at a segment of Rickettsia rickettsii rRNA amplified with polymerase chain reaction. J Clin Microbiol. 1989 Dec;27(12):2692–6.
Wilson, K. H., et al. “Probe directed at a segment of Rickettsia rickettsii rRNA amplified with polymerase chain reaction.J Clin Microbiol, vol. 27, no. 12, Dec. 1989, pp. 2692–96. Pubmed, doi:10.1128/jcm.27.12.2692-2696.1989.
Wilson KH, Blitchington R, Shah P, McDonald G, Gilmore RD, Mallavia LP. Probe directed at a segment of Rickettsia rickettsii rRNA amplified with polymerase chain reaction. J Clin Microbiol. 1989 Dec;27(12):2692–2696.

Published In

J Clin Microbiol

DOI

ISSN

0095-1137

Publication Date

December 1989

Volume

27

Issue

12

Start / End Page

2692 / 2696

Location

United States

Related Subject Headings

  • Staphylococcus aureus
  • Rickettsia typhi
  • Rickettsia rickettsii
  • Rickettsia prowazekii
  • Rickettsia
  • RNA, Viral
  • RNA, Ribosomal, 16S
  • RNA, Ribosomal
  • Polymerase Chain Reaction
  • Oligonucleotide Probes