Unit of Diabetes and Celiac Disease, Department of Clinical Sciences, University Hospital MAS, Lund University, Malmö, Sweden.
Scand J Clin Lab Invest. 2011 Dec;71(8):701-4. doi: 10.3109/00365513.2011.619564. Epub 2011 Oct 3.
Tissue transglutaminase (tTG) autoantibodies are serological markers for celiac disease. The aim was to study the efficacy of the pTnT-rhtTG plasmid and subsequent diagnostic accuracy of tTG autoantibodies for childhood celiac disease using radioligand binding assays.
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.
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.
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.
组织转谷氨酰胺酶（tTG）自身抗体是乳糜泻的血清学标志物。本研究旨在通过放射性配体结合试验，研究 pTnT-rhtTG 质粒的疗效以及 tTG 自身抗体对儿童乳糜泻的诊断准确性。
使用 TNT SP6 耦合网织红细胞裂解物系统，在 [³⁵S]甲硫氨酸存在的情况下，通过 pTnT-rhtTG 以及 pGEMt Easy-rhtTG 载体进行 tTG 的体外转录和翻译。使用两种独立的放射性配体结合试验，分别使用抗人 IgA 琼脂糖和蛋白 A 琼脂糖珠来检测 IgA-tTG 和 IgG-tTG，检测 190 例乳糜泻患儿和 74 例对照者血清中的 tTG 自身抗体。
与 pGEMt Easy-rhtTG 质粒相比，pTnT-rhtTG 中 [³⁵S]甲硫氨酸的掺入中位数为 26%（p = 0.0016）。使用 pTnT-rhtTG（与 pGEMt Easy-rhtTG 相比），IgA-tTG 的敏感性为 96.3%（95.7%）和 IgG-tTG 的敏感性为 95.8%（97.3%），特异性分别为 IgA-tTG = 91.9%（90.5%）和 IgG-tTG = 94.6%（98.4%）。根据 pTnT（pGEMt Easy）试验的受试者工作特征曲线，曲线下面积分别为 IgA-tTG = 98.4%（98.4%）和 IgG-tTG = 97.7%（97.2%）。
pTnT-rhtTG 质粒增加了 tTG 抗原的使用效果，而不降低 IgA-tTG 和 IgG-tTG 对儿童乳糜泻的诊断准确性。因此，建议使用 pTnT-rhtTG 质粒代替 pGEMt Easy-rhtTG 质粒，用于使用放射性配体结合试验评估 IgA-tTG 和 IgG-tTG。
