Comparison of the mismatch-specific endonuclease method and denaturing high-performance liquid chromatography for the identification of HBB gene mutations
Journal
BMC Biotechnology
Journal Volume
8
Date Issued
2008
Author(s)
Hung C.-C.
Su Y.-N.
Lin C.-Y.
Chang Y.-F.
Chang C.-H.
Abstract
Background: Beta-thalassemia is a common autosomal recessive hereditary disease in the Meditertanean, Asia and African areas. Over 600 mutations have been described in the beta-globin (HBB), of which more than 200 are associated with a beta-thalassemia phenotype. Results: We used two highly-specific mutation screening methods, mismatch-specific endonuclease and denaturing high-performance liquid chromatography, to identify mutations in the HBB gene. The sensitivity and specificity of these two methods were compared. We successfully distinguished mutations in the HBB gene by the mismatch-specific endonuclease method without need for further assay. This technique had 100% sensitivity and specificity for the study sample. Conclusion: Compared to the DHPLC approach, the mismatch-specific endonuclease method allows mutational screening of a large number of samples because of its speed, sensitivity and adaptability to semi-automated systems. These findings demonstrate the feasibility of using the mismatch-specific endonuclease method as a tool for mutation screening. ? 2008 Hung et al; licensee BioMed Central Ltd.
SDGs
Other Subjects
Body fluids; Chromatography; Mismatch-specific endonuclease; Liquid chromatography; beta globin; endonuclease; globin; analytic method; article; beta thalassemia; clinical article; controlled study; denaturing high performance liquid chromatography; feasibility study; frameshift mutation; genetic screening; human; intermethod comparison; mismatch specific endonuclease method; missense mutation; mutational analysis; nonsense mutation; nucleotide sequence; sensitivity and specificity; single nucleotide polymorphism; base mispairing; beta thalassemia; comparative study; genetics; genotype; heteroduplex analysis; high performance liquid chromatography; methodology; molecular genetics; mutation; polymerase chain reaction; Base Pair Mismatch; Base Sequence; beta-Thalassemia; Chromatography, High Pressure Liquid; DNA Mutational Analysis; Endonucleases; Genetic Screening; Genotype; Globins; Heteroduplex Analysis; Humans; Molecular Sequence Data; Mutation; Polymerase Chain Reaction; Sensitivity and Specificity
Type
journal article