The use of Taqman genotyping assays for rapid confirmation of β‐thalassaemia mutations in the Malays: accurate diagnosis with low <scp>DNA</scp> concentrations

<jats:title>Summary</jats:title><jats:sec><jats:title>Introduction</jats:title><jats:p>In Malaysia, β‐thalassaemia is a common inherited blood disorder in haemoglobin synthesis with a carrier rate of 4.5%. Currently, <jats:styled-content style="fixed-case">PCR</jats:styled-content>‐incorporating techniques such as amplification refractory mutation system (<jats:styled-content style="fixed-case">ARMS</jats:styled-content>) or reverse dot blot hybridization (<jats:styled-content style="fixed-case">RDBH</jats:styled-content>) are used in β‐thalassaemia mutation detection. <jats:styled-content style="fixed-case">ARMS</jats:styled-content> allows single‐mutation identification using two reactions, one for wild type and another for mutant alleles. <jats:styled-content style="fixed-case">RDBH</jats:styled-content> requires probe immobilization and optimization of hybridization and washing temperatures which is time consuming. The aim of our study was to investigate whether β‐thalassaemia mutations can be identified in samples with low <jats:styled-content style="fixed-case">DNA</jats:styled-content> concentrations.</jats:p></jats:sec><jats:sec><jats:title>Methods</jats:title><jats:p>Genotype identification of common β‐thalassaemia mutations in Malays was carried out using Taqman genotyping assays.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Results show that the Taqman assays allow mutation detection with <jats:styled-content style="fixed-case">DNA</jats:styled-content> template concentrations as low as 2–100 ng. In addition, consistent reproducibility was observed in the Taqman assays when repeated eight times and at different time intervals.</jats:p></jats:sec><jats:sec><jats:title>Conclusion</jats:title><jats:p>The developed sensitive Taqman assays allow molecular characterization of β‐thalassaemia mutations in samples with low <jats:styled-content style="fixed-case">DNA</jats:styled-content> concentrations. The Taqman genotyping assays have potential as a diagnostic tool for foetal blood, chorionic villi or pre‐implantation genetic diagnosis where <jats:styled-content style="fixed-case">DNA</jats:styled-content> is limited and precious.</jats:p></jats:sec>