Comparative performance of ELISA and dot blot assay for TSH-receptor antibody detection in Graves' disease

格雷夫斯病中 TSH 受体抗体检测的 ELISA 和斑点印迹法的比较性能

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作者:Zulkarnain Zulkarnain, Zulvikar Syambani Ulhaq, Hidayat Sujuti, Djoko Wahono Soeatmadji, Hendra Zufry, Dyah Kinasih Wuragil, Agung Pramana W Marhendra, Wibi Riawan, Siti Kurniawati, Yudit Oktanella, Aulanni'am Aulanni'am

Background

Graves' disease (GD) is an autoimmune disease, and it accounts for major cases of hyperthyroidism. Antibody against thyroid-stimulating hormone receptor/TSHR (TRAb) is responsible for hyperthyroidism and is considered as a diagnostic marker for GD. Therefore, we developed a recombinant protein of human TSHR-169 (hTSHR-169), which was specifically recognized TRAb in the serum of GD patients and then compare the diagnostic performance between ELISA and dot blot of TRAb tests for their ability to diagnose GD.

Conclusion

Although the dot blot assay exhibited lower performance than the ELISA method, the dot blot assay is a simple and rapid diagnostic assay that is suitable for diagnosing GD in rural areas, in which healthcare facilities sometimes are not accessible.

Methods

20 GD patients and 20 healthy individuals from the Indonesian population were enrolled. TRAb concentration and density were quantified. Comparative analysis was performed using receiver-operating curve (ROC) analysis.

Results

For dot blot assay, the minimum concentration to detect TRAb requiring 100 ng of antigen with antiserum diluted at 1:60. For diagnosing GD, the ELISA yielded a higher AUC compared with the dot blot assay (0.95 and 0.85, respectively). Using the recommended cutoff values, the efficiency of both assays was examined by comparing the specificity and sensitivity of the assays to the clinical diagnosis. The ELISA showed 80% and 95%, while the dot blot assay showed 70% and 95% sensitivity and specificity, respectively.

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