A SIMPLE AND HIGHLY SENSITIVE ELISA FOR SCREENING OF THE α-THALASSEMIA-1 SOUTHEAST ASIAN-TYPE DELETION

Abstract

Couples in which both partners carry the α-thalassemia-1 trait have a 25% risk of hemoglobin Bart's hydrops fetalis in each pregnancy. Identification of α-thalassemia-1 trait is, therefore, necessary in order to control this severe form of α-thalassemia. We have generated monoclonal antibodies specific to the ζ-globin chain without cross reaction with other globin chains. A simple and sensitive ELISA was developed by using poly-l-lysine to increase the protein binding to the ELISA plate. The developed poly-l-lysine pre-coated ELISA has a very high sensitivity (100%) and specificity (97%) for detection of carriers of α-thalassemia-1 with Southeast Asian-type deletion.

Keywords:

• α-thalassemia-1 screening
• α-thalassemia-1 SEA type
• ζ-globin chain
• Hemoglobin Bart's hydrops fetalis
• monoclonal antibodies
• poly-l-lysine pre-coated ELISA

ACKNOWLEDGMENTS

This article was supported by the NSTDA Research Chair Grant, National Sciences and Technology Development Agency (Thailand), and the National Research University Project under Thailand Office of the Higher Education Commission. We thank Anusorn Lekawipat and Kantinan Chuensirikulchai for the technical assistances. Saichit Khummuang is a doctoral candidate of the Biomedical Sciences program, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand. We thank Dr. Dale E. Taneyhill for proofreading the manuscript.

Notes

The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), false positive rate, and accuracy of the test are 100%, 96.67%, 96.67%, 96.67%, 3.33%, and 98.33%, respectively.