Detection of alpha-thalassemia-1 Southeast Asian type using real-time gap-PCR with SYBR Green1 and high resolution melting analysis
Detection of alpha-thalassemia-1 Southeast Asian type using real-time gap-PCR with SYBR Green1 and high resolution melting analysis
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DOI：
10.1111/j.1600-0609.2008.01055.x
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发表时间：
2008-06-01
影响因子：
3.1
通讯作者：
Sanguansermsri, Torpong
中科院分区：
文献类型：
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作者：
Pornprasert, Sakorn;Phusua, Arunee;Sanguansermsri, Torpong
α -地中海贫血-1型东南亚（SEA）型是亚洲人群中最常见的遗传性疾病。双方都是携带者的夫妇有25%的机会怀上巴特氏水肿胎儿。因此，经常需要快速获得携带者筛查和产前诊断的结果。建立了一种快速诊断-地中海贫血-1 SEA型的方法。所使用的技术是基于实时间隙pcr和使用Rotor-Gene 6000 （TM）对扩增片段进行高分辨率熔化（HRM）分析。用于扩增的DNA样本来自全血、脐带血和绒毛膜绒毛取样（CVS）。用这种方法可以很容易地将α -地中海贫血-1 SEA等位基因与野生型α -珠蛋白基因等位基因区分开来。实时gap-PCR和HRM分析提供了额外的好处，包括最少的劳动力，快速的周转时间，并降低了PCR遗留污染的风险。它具有成本效益和安全性，不需要荧光标记探针和危险化学品。此外，它是准确的显示100%的一致性与传统的gap-PCR分析。
alpha-Thalassemia-1 Southeast Asian (SEA) type is the most common genetic disorder in the Asian population. Couples who are both carriers have a 25% chance of conceiving Bart's hydrops fetalis. Therefore, results from carrier screening and prenatal diagnosis frequently need to be available rapidly. A rapid technique for diagnosis of alpha-thalassemia-1 SEA type was implemented. The technique used is based on real-time gap-PCR and high resolution melting (HRM) analysis of the amplified fragment using the Rotor-Gene 6000 (TM). The DNA samples used for amplification were obtained from whole blood, cord blood, and chorionic villus sampling (CVS). With this method, the alpha-thalassemia-1 SEA allele can be easily distinguished from wild type alpha-globin gene allele. The real-time gap-PCR and HRM analysis offers additional benefits including minimal labor, rapid turnaround time, and a decreased risk of PCR carryover contamination. It is cost-effective and safe, does not require fluorescently labeled probe and hazardous chemicals. Moreover, it is accurate showing 100% concordance with conventional gap-PCR analysis.