Rapid detection of α-thalassaemia alleles of --SEA/, -α3.7/ and -α4.2/ using a dual labelling, self-quenching hybridization probe/melting curve analysis. Issue 6 (December 2015)

• Record Type: Journal Article
• Authors: Gao, Lan; Liu, Yanhui; Sun, Manna; Zhao, Ying; Xie, Rungui; He, Yi; Xu, Wanfang; Liu, Jianxin; Lin, Yangyang; Lou, Jiwu
• Is Part Of: Molecular and cellular probes. Volume 29:Issue 6(2015)
• Page Start: 438
• Page End: 441
• Publication Date: 2015-12
• Subjects: Melting curve -- Dually labelled probes -- α-Thalassaemia -- Deletion; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2015.07.004
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: Objectives: The aim of the study was to set up an alternative automatic molecular diagnostic method for deletional α-thalassaemia mutations without gel electrophoresis. Methods: Based on the sequence variation within the two Z boxes and melting curve analysis of dually labelled probes, a real-time PCR assay was developed and validated for the rapid detection of major α-genotypes (-- SEA /αα, -- SEA /-α 3.7, -- SEA /-α 4.2, -- SEA /-- SEA, -α 3.7 /-α 3.7 and -α 4.2 /-α 4.2 ). Results: Samples with the -α 3.7 /-α 3.7, -α 4.2 /-α 4.2, -- SEA /αα, -- SEA /-α 3.7, -- SEA /-α 4.2, and -- SEA /-- SEA genotypes could be clearly distinguished. The accuracy of this technique for these samples was 100% sensitivity and specificity. Conclusion: This technique is rapid and reliable, demonstrating feasibility for use in large-scale population screening and prenatal diagnosis of deletional Hb H disease and Hb Bart's hydrops fetalis. Highlights: We developed a rapid and reliable real-time PCR/melting curve analysis assay to rapidly detect the major α-genotypes. The -α 3.7 /-α 3.7, -α 4.2 /-α 4.2, -- SEA /αα, -- SEA /-α 3.7, -- SEA /-α 4.2 and -- SEA /-- SEA genotypes could be detected. The accuracy of this technique for our sample was 100% sensitivity and specificity.