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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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Abstract

α‐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 α‐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™. The DNA samples used for amplification were obtained from whole blood, cord blood, and chorionic villus sampling (CVS). With this method, the α‐thalassemia‐1 SEA allele can be easily distinguished from wild type α‐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.
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Document Type: Research Article

Affiliations: 1: Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang-Mai University, Chiang-Mai, Thailand 2: Division of Hematology, Department of Pediatrics, Faculty of Medicine, Chiang-Mai University, Chiang-Mai, Thailand

Publication date: June 1, 2008

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