Detection of alpha-thalassemia-1 Southeast Asian type using real-time gap-PCR with SYBR Green1 and high resolution melting analysis

• Main Authors: Sakorn Pornprasert, Arunee Phusua, Sudjai Suanta, Rattika Saetung, Torpong Sanguansermsri
• Format: Journal
• Published: 2018
• Subjects: Medicine
• Online Access: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=43849098381&origin=inward; http://cmuir.cmu.ac.th/jspui/handle/6653943832/60624
• Institution: Chiang Mai University

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 hazardoucs chemicals. Moreover, it is accurate showing 100% concordance with conventional gap-PCR analysis. © 2008 The Authors.