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• W3045112179 abstract "Abstract Introduction Several DNA‐based approaches including a reverse dot‐blot hybridization (RDB) have been established for detection of β‐thalassemia genotypes to provide accurate genetic counseling and prenatal diagnosis for prevention and control of severe β‐thalassemia. However, one of major concerns of these techniques is a risk of misdiagnosis due to a lack of DNA controls. Here, we constructed positive DNA controls for β‐thalassemia genotyping in order to ensure that all steps in the analysis are performed properly. Methods Four recombinant β‐globin plasmids, including a normal sequence and three different mutant panels covering 10 common β‐thalassemia mutations in Asia, were constructed by a conventional cloning method followed by sequential rounds of site‐directed mutagenesis. These positive DNA controls were further validated by RDB analysis. Results We demonstrated the applicability of established positive DNA controls for β‐thalassemia genotyping in terms of accuracy and reproducibility by RDB analysis. We further combined three mutant β‐globin plasmids into a single positive control, which showed positive signals for both normal and mutant probes of all tested mutations. Therefore, only two positive DNA controls, normal and combined mutant β‐globin plasmids, are required for detecting 10 common β‐thalassemia mutations by RDB, reducing the cost, time, and efforts in the routine diagnosis. Conclusion The β‐globin DNA controls established here provide efficient alternatives to a conventional DNA source from peripheral blood, which is more difficult to obtain. They also provide a platform for future development of β‐globin plasmid controls with other mutations, which can also be suitable for other DNA‐based approaches." @default.
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• W3045112179 date "2020-07-24" @default.
• W3045112179 modified "2023-09-27" @default.
• W3045112179 title "Development of DNA controls for detection of β‐thalassemia mutations commonly found in Asian" @default.
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• W3045112179 doi "https://doi.org/10.1111/ijlh.13292" @default.
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