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首页> 外文期刊>Annals of laboratory medicine. >Detection of α-thalassemia-1 Southeast Asian and Thai Type Deletions and β-thalassemia 3.5-kb Deletion by Single-tube Multiplex Real-time PCR with SYBR Green1 and High-resolution Melting Analysis
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Detection of α-thalassemia-1 Southeast Asian and Thai Type Deletions and β-thalassemia 3.5-kb Deletion by Single-tube Multiplex Real-time PCR with SYBR Green1 and High-resolution Melting Analysis

机译：SYBR Green1单管多重实时PCR和高分辨率熔解分析法检测α-地中海贫血-1东南亚和泰国型缺失和β-地中海贫血3.5-kb缺失

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Background: Prevention and control of thalassemia requires simple, rapid, and accurate screening tests for carrier couples who are at risk of conceiving fetuses with severe thalassemia. Methods: Single-tube multiplex real-time PCR with SYBR Green1 and high-resolution melting (HRM) analysis were used for the identification of α-thalassemia-1 Southeast Asian (SEA) and Thai type deletions and β-thalassemia 3.5-kb gene deletion. The results were compared with those obtained using conventional gap-PCR. DNA samples were derived from 28 normal individuals, 11 individuals with α-thalassemia-1 SEA type deletion, 2 with α-thalassemia-1 Thai type deletion, and 2 with heterozygous β-thalassemia 3.5-kb gene deletion. Results: HRM analysis indicated that the amplified fragments from α-thalassemia-1 SEA type deletion, α-thalassemia-1 Thai type deletion, β-thalassemia 3.5-kb gene deletion, and the wild-type β-globin gene had specific peak heights at mean melting temperature (Tm) values of 86.89?C, 85.66?C, 77.24?C, and 74.92?C, respectively. The results obtained using single-tube multiplex real-time PCR with SYBR Green1 and HRM analysis showed 100% consistency with those obtained using conventional gap-PCR. Conclusions: Single-tube multiplex real-time PCR with SYBR Green1 and HRM analysis is a potential alternative for routine clinical screening of the common types of α- and β-thalassemia large gene deletions, since it is simple, cost-effective, and highly accurate.
机译：背景：地中海贫血的预防和控制要求对携带严重地中海贫血胎儿风险的携带者夫妇进行简单，快速和准确的筛查测试。方法：采用SYBR Green1单管多重实时荧光定量PCR和高分辨率熔解（HRM）分析技术鉴定α-地中海贫血-1东南亚（SEA）和泰国型缺失以及β-地中海贫血3.5-kb基因删除。将结果与使用常规gap-PCR获得的结果进行比较。 DNA样本来自28个正常个体，11个具有α-地中海贫血1 SEA型缺失，2个具有α-地中海贫血1 Thai类型缺失和2个具有杂合性β-地中海贫血3.5kb基因缺失的个体。结果：HRM分析表明，α-地中海贫血-1 SEA型缺失，α-地中海贫血-1 Thai类型缺失，β-地中海贫血3.5-kb基因缺失和野生型β-珠蛋白基因的扩增片段具有特定的峰高在平均熔化温度（Tm）下的温度分别为86.89℃，85.66℃，77.24℃和74.92℃。使用具有SYBR Green1的单管多重实时PCR和HRM分析获得的结果显示，与使用常规gap-PCR获得的结果具有100％的一致性。结论：SYBR Green1和HRM分析的单管多重实时PCR是常规临床筛查常见的α型和β型地中海贫血大基因缺失类型的潜在选择，因为它简单，经济高效且高度准确。

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