-
Immunostick Test for Detecting zeta-Globin Chains and Screening of the Southeast Asian alpha-Thalassemia 1 Deletion
- Back
Document Title
Immunostick Test for Detecting zeta-Globin Chains and Screening of the Southeast Asian alpha-Thalassemia 1 Deletion
Author
Pata S, Pongpaiboon M, Laopajon W, Munkongdee T, Paiboonsukwong K, Pornpresert S, Fucharoen S, Kasinrerk W
Name from Authors Collection
Affiliations
Chiang Mai University; Chiang Mai University; National Science & Technology Development Agency - Thailand; National Center Genetic Engineering & Biotechnology (BIOTEC); Mahidol University; Chiang Mai University
Type
Article
Source Title
BIOLOGICAL PROCEDURES ONLINE
Year
2019
Volume
21
Page
-
Open Access
Green Published, gold
Publisher
BMC
DOI
10.1186/s12575-019-0104-2
Format
Abstract
BackgroundCouples who carry alpha-thalassemia-1 deletion are at 25% risk of having a fetus with hemoglobin Bart's hydrops fetalis. Southeast Asian deletion (--(SEA)) is the most common type of alpha-thalassemia 1 among Southeast Asian populations. Thus, identification of the (--(SEA)) alpha-thalassemia 1 carrier is necessary for controlling severe alpha-thalassemia in Southeast Asian countries.ResultsUsing our generated anti zeta-globin chain monoclonal antibodies (mAbs) clones PL2 and PL3, a simple immunostick test for detecting zeta-globin chain presence in whole blood lysates was developed. The procedure of the developed immunostick test was as follows. The immunostick paddles were coated with 50 mu g/mL of mAb PL2 as capture mAb, or other control antibodies. The coated immunostick was dipped into cocktail containing tested hemolysate at dilution of 1:500, 0.25 mu g/mL biotin-labeled mAb PL3 and horseradish peroxidase-conjugated streptavidin at dilution of 1:1000. The immunostick was then dipped in precipitating substrate and the presence of zeta-globin chain in the tested sample was observed by the naked eye. Upon validation of the developed immunostick test with various types of thalassemia and normal subjects, 100% sensitivity and 82% specificity for detection of the (--(SEA)) alpha-thalassemia-1 carriers were achieved. The mAb pre-coated immunostick can be stored at room temperature for at least 20weeks.ConclusionIn this study, a novel simple immunostick test for the screening of (--(SEA)) alpha-thalassemia 1 carriers was presented. The developed immunostick test, within a single test, contains both positive and negative internal procedural controls.
Funding Sponsor
TRF [RTA5980007]; Thailand Office of Higher Education Commission [MRG6080269, MRG6180253]; TRF Distinguished Research Professor Grant [DPG6080003]; Chiang Mai University (Chiang Mai Center of Excellence Project); Innovation Hubs of Thailand 4.0 project (Chiang Mai University); Innovation Hubs of Thailand 4.0 project (Mahidol University)
Publication Source
WOS