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  • Title: Development and validation of a zeta-globin-specific ELISA for carrier screening of the (--SEA) alpha thalassaemia deletion.
    Author: Tang L, Zhu P, Zhou WJ, Zheng J, Zhou YQ, Fu N, Xu XM.
    Journal: J Clin Pathol; 2009 Feb; 62(2):147-51. PubMed ID: 19181632.
    Abstract:
    AIMS: The Southeast Asian (SEA) deletion (--(SEA)) represents the most common determinant causing alpha thalassaemia in Southeast Asian countries. The embryonic zeta-globin chain has been defined as a marker for the detection of this deletion in adults. The aim of this study was to develop an appropriate low-cost ELISA for zeta-globin chain detection as a routine screening test for (--(SEA)) alpha thalassaemia deletion. METHODS: A sandwich ELISA system for zeta-globin chains was established with a pair of zeta-globin-specific monoclonal antibodies prepared in-house, and locally made products. Against a gap-PCR method that was taken as the standard, this assay was validated in a cohort study testing a total of 526 individuals comprising patients scheduled for haemoglobinopathy diagnostic analysis and normal individuals. Routine screening of the (--(SEA)) deletion in 300 random student volunteers was conducted using the assay. RESULTS: While the cut-off point was set at a percentage positive value of 30, the sensitivity and specificity of this ELISA method were 100% and 99.24%, respectively. The mean intra-assay and inter-assay coefficients of variation among the different concentrations in the optimised ELISA conditions were 2.1-11.4% and 4.3-13.2%, respectively. Seventeen of the 300 volunteers sampled were determined by the ELISA to have the (--(SEA)) deletion; these results were in 100% agreement with the gap-PCR results. CONCLUSIONS: This study validates the ELISA method described here as a simple, rapid and cost-effective assay that is potentially adaptable for application in large-scale population screening for this prevalent disorder in SEA areas such as southern China.
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