Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis

N. S. Sato, T. Suzuki, Takuya Ueda, K. Watanabe, R. D.C. Hirata, M. H. Hirata

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Three recombinant antigens of Treponema pallidum Nichols strain were fused with GST, cloned and expressed in Escherichia coli, resulting in high levels of GST-rTp47 and GST-rTp17 expression, and supplementation with arginine tRNA for the AGR codon was needed to obtain GST-rTp15 overexpression. Purified fusion protein yields were 1.9, 1.7 and 5.3 mg/l of cell culture for GST-rTp47, GST-rTp17 and GST-rTp15, respectively. The identities of the antigens obtained were confirmed by automated DNA sequencing using ABI Prism 310 and peptide mapping by Finningan LC/MS. These recombinant antigens were evaluated by immuno-slot blot techniques applied to 137 serum samples from patients with a clinical and laboratory diagnosis of syphilis (61 samples), from healthy blood donors (50 samples), individuals with sexually transmitted disease other than syphilis (3 samples), and from individuals with other spirochetal diseases such as Lyme disease (20 samples) and leptospirosis (3 samples). The assay had sensitivity of 95.1% (95% CI, 86.1 to 98.7%) and a specificity of 94.7% (95% CI, 87.0 to 98.7%); a stronger reactivity was observed with fraction rTp17. The immunoreactivity results showed that fusion recombinant antigens based-immuno-slot blot techniques are suitable for use in diagnostic assays for syphilis.

Original languageEnglish
Pages (from-to)949-955
Number of pages7
JournalBrazilian Journal of Medical and Biological Research
Volume37
Issue number7
DOIs
Publication statusPublished - 2004 Jan 1
Externally publishedYes

Fingerprint

Syphilis Serodiagnosis
Syphilis
Antigens
Assays
Fusion reactions
Treponema pallidum
Leptospirosis
Peptide Mapping
Lyme Disease
Clinical Laboratory Techniques
Sexually Transmitted Diseases
Transfer RNA
Prisms
Blood Donors
DNA Sequence Analysis
Cell culture
Codon
Escherichia coli
Arginine
Blood

Keywords

  • Cloning
  • GST
  • Recombinant antigen
  • Syphilis
  • Treponema pallidum

ASJC Scopus subject areas

  • Biophysics
  • Neuroscience(all)
  • Biochemistry
  • Physiology
  • Immunology
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Cell Biology

Cite this

Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis. / Sato, N. S.; Suzuki, T.; Ueda, Takuya; Watanabe, K.; Hirata, R. D.C.; Hirata, M. H.

In: Brazilian Journal of Medical and Biological Research, Vol. 37, No. 7, 01.01.2004, p. 949-955.

Research output: Contribution to journalArticle

Sato, N. S. ; Suzuki, T. ; Ueda, Takuya ; Watanabe, K. ; Hirata, R. D.C. ; Hirata, M. H. / Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis. In: Brazilian Journal of Medical and Biological Research. 2004 ; Vol. 37, No. 7. pp. 949-955.
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