Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis
Braz. j. med. biol. res
;
37(7): 949-955, July 2004. ilus, tab
Article
in English
| LILACS
| ID: lil-360935
RESUMO
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 percent (95 percent CI, 86.1 to 98.7 percent) and a specificity of 94.7 percent (95 percent CI, 87.0 to 98.7 percent); 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.
Full text:
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Index:
LILACS (Americas)
Main subject:
Treponema pallidum
/
Recombinant Fusion Proteins
/
Syphilis Serodiagnosis
/
Syphilis
/
Antigens, Bacterial
Type of study:
Diagnostic study
/
Observational study
Limits:
Humans
Language:
English
Journal:
Braz. j. med. biol. res
Journal subject:
Biology
/
Medicine
Year:
2004
Type:
Article
/
Congress and conference
Affiliation country:
Brazil
/
Japan
Institution/Affiliation country:
Instituto Adolfo Lutz/BR
/
Universidade de São Paulo/BR
/
University of Tokyo/JP
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