High resolution melting analytical platform for rapid prenatal and postnatal diagnosis of β-thalassemia common among Southeast Asian population

• We have applied the HRM analysis to diagnosis of β-thalassemia commonly found in Southeast Asian populations.
• Apart from those with DNA deletions, the results of HRM assay matched 100% with routine diagnosis made by other methods.
• The HRM assay could also initially recognize unknown mutations on the amplified fragments.
• The assay should provide a high throughput platform for screening of β-thalassemia common in the region.

BackgroundHigh resolution melting (HRM) analysis is a powerful technology for scanning sequence alteration. We have applied this HRM assay to screen common β-thalassemia mutations found among Southeast Asian population.MethodsKnown DNA samples with 8 common mutations were used in initial development of the methods including − 28 A–G, codon 17 A–T, IVSI-1 G–T, IVSI-5 G–C, codon 26 G–A (Hb E), codons 41/42 − TTCT, codons 71/72 + A and IVSII-654 C–T. Further validation was done on 60 postnatal and 6 prenatal diagnoses of β-thalassemia.ResultsEach mutation has specific HRM profile which could be used in rapid screening. Apart from those with DNA deletions, the results of HRM assay matched 100% with those of routine diagnosis made by routine allele specific PCR. In addition, the HRM assay could initially recognize three unknown mutations including a hitherto un-described one in Thai population.ConclusionsThe established HRM assay should prove useful for rapid and high throughput platform for screening and prenatal diagnosis of β-thalassemia common in the region.