Diagnosis of α-thalassaemia by colorimetric gap loop mediated isothermal amplification
5
Issued Date
2023-12-01
Resource Type
eISSN
20452322
Scopus ID
2-s2.0-85161862128
Pubmed ID
37311778
Journal Title
Scientific Reports
Volume
13
Issue
1
Rights Holder(s)
SCOPUS
Bibliographic Citation
Scientific Reports Vol.13 No.1 (2023)
Suggested Citation
Chumworathayee W., Munkongdee T., Buasuwan N., Chaichompoo P., Svasti S. Diagnosis of α-thalassaemia by colorimetric gap loop mediated isothermal amplification. Scientific Reports Vol.13 No.1 (2023). doi:10.1038/s41598-023-36676-2 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/87635
Title
Diagnosis of α-thalassaemia by colorimetric gap loop mediated isothermal amplification
Other Contributor(s)
Abstract
α-Thalassaemia is an inherited haemoglobin disorder that results from the defective synthesis of α-globin protein. Couples whom both carry the α-thalassaemia 1 gene are at risk of having a foetus with the most severe thalassaemia, Hb Bart’s hydrops fetalis, with a risk of maternal mortality. However, haematological parameters alone cannot distinguish between a α-thalassaemia 1 carrier and a homozygous α-thalassaemia 2, in which one α-globin gene has been deleted on each chromosome. A rapid and accurate molecular detection assay is essential for prevention of the disease in populations where α-thalassaemia 1 is common. Multiplex Gap-PCR analysis is widely used for diagnosis of α-thalassaemia. However, the technique requires a thermocycler and post-amplification processing, which limits its application in primary care or in rural areas in developing countries. Loop mediated isothermal amplification (LAMP) amplifies target DNA at a constant temperature and does not require a thermocycler. This study developed a colorimetric Gap-LAMP using malachite green to allow naked eye visualization of two deletional α-thalassaemia 1 commonly found in Asian populations, the Southeast Asian type (--SEA) and the Thai type (--THAI) deletions. The Gap-LAMP was performed on DNA samples from 410 individuals carrying various α-thalassaemia gene defects with 100% concordance with conventional Gap-PCR analysis. This method eliminates post-amplification processing or the use of expensive sophisticated equipment and allows screening large populations for the prevention and control of α-thalassaemia.