Comparison of the mismatch-specific endonuclease method and denaturing high-performance liquid chromatography for the identification of HBB gene mutations.
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.


CONCLUSIONS
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.
Hung
Chia-Cheng
CC
1Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan. chiacheng@ntu.edu.tw
Su
Yi-Ning
YN
Lin
Chia-Yun
CY
Chang
Yin-Fei
YF
Chang
Chien-Hui
CH
Cheng
Wen-Fang
WF
Chen
Chi-An
CA
Lee
Chien-Nan
CN
Lin
Win-Li
WL
D020137:Base Pair Mismatch; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004252:DNA Mutational Analysis / Q000379:methods; D004720:Endonucleases / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods; D005838:Genotype; D005914:Globins / Q000235:genetics*; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017086:beta-Thalassemia / Q000235:genetics
D005914:Globins; D004720:Endonucleases