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[Quality control of radioligand assay for glutamic acid decarboxylase antibody]
Gan Huang1, Zhi-guang Zhou, Jian Peng
1Institute of Metabolism and Endocrinology, Xiangya Second Hospital, Central South University, Changsha 410011, China.
Objective:
To supply the quality control for glutamic acid decarboxylase antibody (GAD-Ab) measurement using radioligand assay.
Methods:
The quality control of GAD-Ab determined with radioligand assay in 83 batches was analyzed.
Results:
The intra-assay coefficient of variation (CV) was 4.3% to approximately 8.3% and the inter-assay CV was 7.1% to approximately 10.8%. A ratio of signal to noise (S/N) with 15 or more was defined as a valid measurement. Of the 83 batches, 67 batches (80.7%) met with the standard and the other 16 (19.3%) were not qualified. When the results were analyzed according to the season, the failure rate in summer was significantly higher than that in winter (38.5% vs 4.8%, P < 0.05). In addition, the mean value of S/N in summer was significantly lower than that in winter ( 14.1 +/- 7. 3 vs. 21.5 +/- 5.2, P < 0.01). Among the failed batches, 68.8% (11/16) was caused by TBST buffer and 50% (8/16) due to the increased temperature of TBST buffer. Other causes for the failure included the invalidation of reagents for in vitro transcription and translation.
Conclusion:
TBST buffer and other influential factors of radioligand assay in determining GAD-Ab, should be controlled strictly to ensure a reliable and repeatable result.
