Efficiency and cost effectiveness: PAGE-SSCP versus MDE and Phast gels for the identification of unknown beta thalassaemia mutations.

A Gupta, S Agarwal
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引用次数: 14

Abstract

Background: Prenatal diagnosis for β thalassaemia has proved to be very effective in preventing the birth of an affected child and hence in controlling the disease. The success of prenatal diagnosis depends on the delineation of the underlying mutations in the population at risk. Each population carries a limited number of frequent defects (89–91%) and a variable number of rare alleles (4–5%), whereas 2–3% of alleles remain uncharacterised. To offer prenatal diagnosis when the parental mutation is unknown, the application of a non-specific detection method (such as single stranded conformational polymorphism (SSCP)) to localise the mutation, followed by direct sequencing of the amplified gene sequence, is required. With this objective in mind, this study was designed to devise the best protocol and system of SSCP for the rapid screening of unknown mutations in the β globin gene. Methods: To detect mutations in this disease, three different systems—Phast gels, MDE gels, and polyacrylamide gels—were used under varying conditions. Results: Polyacrylamide gels were found to be the most efficient, both in terms of resolution and cost. Conclusion: Polyacrylamide gels are the most rapid, efficient, reliable, and cost effective means for DNA mutation analysis of the β globin gene.
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效率和成本效益:PAGE-SSCP与MDE和Phast凝胶鉴别未知地中海贫血突变
背景:对地中海贫血的产前诊断已被证明对预防患病儿童的出生并因此控制该疾病非常有效。产前诊断的成功取决于对高危人群潜在突变的描述。每个人群携带有限数量的常见缺陷(89-91%)和可变数量的罕见等位基因(4-5%),而2-3%的等位基因仍未被表征。为了在亲本突变未知的情况下提供产前诊断,需要应用非特异性检测方法(如单链构象多态性(SSCP))来定位突变,然后对扩增的基因序列进行直接测序。考虑到这一目标,本研究旨在设计最佳的SSCP方案和系统,用于快速筛选β -珠蛋白基因的未知突变。方法:在不同的条件下使用三种不同的系统- phast凝胶,MDE凝胶和聚丙烯酰胺凝胶来检测该疾病的突变。结果:聚丙烯酰胺凝胶在分辨率和成本方面都是最有效的。结论:聚丙烯酰胺凝胶是快速、高效、可靠、经济的β -珠蛋白基因突变分析方法。
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