Abstract | BACKGROUND: METHODS: Coupled in vitro transcription and translation of tTG were performed by pTnT-rhtTG as well as by the pGEMt Easy-rhtTG vectors using the TNT SP6 Coupled Reticulocyte Lysate System in the presence of [³⁵S] methionine. Sera from 190 celiac disease children and 74 controls were measured for tTG autoantibodies in two separate radioligand binding assays using anti-human IgA agarose and protein A sepharose beads for the detection of IgA-tTG and IgG-tTG, respectively. RESULTS: Median incorporation of [³⁵S] methionine into the pTnT-rhtTG was 26% compared to 16% for the pGEMt Easy-rhtTG plasmid (p = 0.0016). Using pTnT-rhtTG (as compared to pGEMt Easy-rhtTG), sensitivities were IgA-tTG = 96.3% (95.7%) and IgG-tTG = 95.8% (97.3%) and specificities were IgA-tTG = 91.9% (90.5%) and IgG-tTG = 94.6% (98.4%). According to receiver operator characteristics for the pTnT (pGEMt Easy) assays, area under the curves were IgA-tTG = 98.4% (98.4%) and IgG-tTG = 97.7% (97.2%), respectively. CONCLUSION: The pTnT-rhtTG plasmid increased the efficacy of tTG antigen usage without reducing the diagnostic accuracy of IgA-tTG and IgG-tTG for childhood celiac disease. The pTnT-rhtTG plasmid is therefore recommended over the pGEMt Easy-rhtTG for the assessment of IgA-tTG and IgG-tTG using radioligand binding assays.
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Authors | Jennifer Kjellerås, Fariba Vaziri-Sani, Daniel Agardh |
Journal | Scandinavian journal of clinical and laboratory investigation
(Scand J Clin Lab Invest)
Vol. 71
Issue 8
Pg. 701-4
(Dec 2011)
ISSN: 1502-7686 [Electronic] England |
PMID | 21961813
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antibodies, Anti-Idiotypic
- Antigens, Neoplasm
- Autoantibodies
- Biomarkers
- Staphylococcal protein A-sepharose
- Sulfur Radioisotopes
- anti-IgA
- Sepharose
- Methionine
- Transglutaminases
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Topics |
- Adolescent
- Antibodies, Anti-Idiotypic
(immunology, metabolism)
- Antigens, Neoplasm
(blood, immunology)
- Autoantibodies
(blood, immunology)
- Biomarkers
(blood)
- Case-Control Studies
- Celiac Disease
(blood, diagnosis, immunology)
- Cell-Free System
(metabolism)
- Child
- Child, Preschool
- Female
- Humans
- Infant
- Male
- Methionine
(analysis, metabolism)
- Plasmids
(chemistry, immunology)
- Protein Binding
- ROC Curve
- Radioligand Assay
- Reticulocytes
(metabolism)
- Sepharose
(analogs & derivatives, metabolism)
- Sulfur Radioisotopes
(analysis, metabolism)
- Sweden
- Transglutaminases
(blood, immunology)
- Young Adult
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