(快速检测的常见alpha-thalassemia-2因素PCR试验)。
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刘赵Y,钟M, Z,徐X
(快速检测的常见alpha-thalassemia-2因素PCR试验)。
易建联中华雪川雪。2001年6月;18 (3):216 - 8。
- PubMed ID
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11402454 (在PubMed]
- 文摘
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目的:最常见的分子病变alpha-thalassemia-2决定因素是一个身体基因的缺失。满意的PCR方法检测删除所需的分子诊断和遗传筛查以来没有内部控制在大多数发表的PCR策略。本研究的目的是开发一个可靠的PCR协议特定的两个常见alpha-thalassemia-2与内部控制因素。方法:身体的多个重复的元素和高GC-content轨迹实施严厉限制设计合适的引物PCR和优化稳定条件。在这项研究中,两种复合PCR系统被成功建立。向右一个旨在检测删除(- alpha(3.7) /)与两对引物包括一对新优化的内部标准的放大显示的成功失败的PCR扩增。,左侧的删除(t1 -(4.2) /)三个引物,是根据新测序的数据设计- alpha(4.2)和HbQ-alpha(4.2)在这个实验室(加入基因库没有删除。AF221717)。PCR体系,一个是作为常见的上游引物和其他两个是用作输入特定的下游引物正常等位基因和删除一个,分别。结果:更容易解释的,明确的放大观察使用多重PCR系统检测的两种常见alpha-thalassemia-2决定因素。 The three or four primers were run in the same tube under the same condition and both of these two systems could be used at the same thermal cycle parameters. For typing the rightward deletion, a mutant-specific amplification of 1.7 kb and a 1140 bp amplified band as a normal and system control were produced. For typing the leftward deletion, two PCR-amplified bands, a 956 bp fragment specific for a -alpha(4.2) gene and a 1140 bp one for a normal allele were found. CONCLUSION: Two sets of PCR systems with internal controls for detecting the most common two alpha-thalassemia-2 determinants have been established and may be suitable for molecular diagnosis and population screening.