High resolution DNA melting analysis: an application for prenatal control of alpha-thalassemia

Prenatal Diagnosis
S SirichotiyakulTorpong Sanguansermsri

Abstract

To report the use of real-time gap-PCR using SYTO9 with high-resolution melting analysis (HRMA) in prenatal diagnosis of alpha-thalassemia 1. Real-time gap-PCR using SYTO9 with HRMA was performed in 33 DNA samples from chorionic villi sampling (8 normal, 16 heterozygous, and 9 homozygous) to determine the alpha-thalassemia 1 gene [normal and Southeast Asia (-SEA) allele]. The dissociation curve analysis in normal and - SEA allele gave a peak of T(m) at 91.80 +/- 0.14 degrees C and 88.67 +/- 0.08 degrees C, respectively. Normal genotype and homozygous alpha-thalassemia 1 showed a single peak of T(m) that corresponded to their alleles. The heterozygotes gave both peaks with higher normal peak and smaller - SEA peak. Thirty one samples showed consistent results with the conventional gap-PCR. Two samples with ambiguous results were confirmed to be maternal DNA contamination on real-time quantitative PCR and microsatellite assay. HRMA from both samples showed similar pattern to that of heterozygotes. However, they showed much smaller normal peak compared with the - SEA peak, which is in contrast to those of heterozygotes and can readily be distinguished. HRMA with SYTO9 is feasible for prenatal diagnosis of alpha-thalassemia. It had...Continue Reading

References

May 22, 2001·Prenatal Diagnosis·M KleanthousM Angastiniotis
Jun 1, 2005·Hemoglobin·Pimlak CharoenkwanTorpong Sanguansermsri
Aug 5, 2006·Translational Research : the Journal of Laboratory and Clinical Medicine·Jingzhong LiuBai Xiao
Feb 12, 2009·Hemoglobin·S SirichotiyakulTorpong Sanguansermsri

Citations

Dec 18, 2013·European Journal of Haematology·Sakorn Pornprasert, Watcharee Prasing

Related Concepts

Bimonthly
DNA, Double-Stranded
Fluorochromes
RNA Denaturation
Organic Chemicals
Chorionic Villi Sampling
Whole Genomic DNA Probes
Nested Polymerase Chain Reaction
Alpha-Thalassemia
Alleles

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