Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays

Beta-thalassemia is a life-threatening inherited blood disorder. Rapid characterization of β-globin gene mutations is necessary because of the high frequency of Malaysian β-thalassemia carriers. A combination real-time polymerase chain reaction genotyping assay using TaqMan probes was developed to c...

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Main Authors: Kho, Siew Leng, Chua, Kek Heng, George, Elizabeth, Tan, Mary Anne Jin Ai
Format: Article
Language:English
Published: Fundacao de Pesquisas Cientificas de Ribeirao Preto 2013
Online Access:http://psasir.upm.edu.my/id/eprint/29670/
http://psasir.upm.edu.my/id/eprint/29670/1/Specific%20and%20straightforward%20molecular%20investigation%20of%20%CE%B2.pdf
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author Kho, Siew Leng
Chua, Kek Heng
George, Elizabeth
Tan, Mary Anne Jin Ai
author_facet Kho, Siew Leng
Chua, Kek Heng
George, Elizabeth
Tan, Mary Anne Jin Ai
author_sort Kho, Siew Leng
building UPM Institutional Repository
collection Online Access
description Beta-thalassemia is a life-threatening inherited blood disorder. Rapid characterization of β-globin gene mutations is necessary because of the high frequency of Malaysian β-thalassemia carriers. A combination real-time polymerase chain reaction genotyping assay using TaqMan probes was developed to confirm β-globin gene mutations. In this study, primers and probes were designed to specifically identify 8 common β-thalassemia mutations in the Malaysian Malay and Chinese ethnic groups using the Primer Express software. "Blind tests" using DNA samples from healthy individuals and β-thalassemia patients with different genotypes were performed to determine the specificity and sensitivity of this newly designed assay. Our results showed 100% sensitivity and specificity for this novel assay. In conclusion, the TaqMan genotyping assay is a straightforward assay that allows detection of β-globin gene mutations in less than 40 min. The simplicity and reproducibility of the TaqMan genotyping assay permit its use in laboratories as a rapid and cost-effective diagnostic tool for confirmation of common β-thalassemia mutations in Malaysia.
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spelling upm-296702016-11-30T02:28:10Z http://psasir.upm.edu.my/id/eprint/29670/ Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays Kho, Siew Leng Chua, Kek Heng George, Elizabeth Tan, Mary Anne Jin Ai Beta-thalassemia is a life-threatening inherited blood disorder. Rapid characterization of β-globin gene mutations is necessary because of the high frequency of Malaysian β-thalassemia carriers. A combination real-time polymerase chain reaction genotyping assay using TaqMan probes was developed to confirm β-globin gene mutations. In this study, primers and probes were designed to specifically identify 8 common β-thalassemia mutations in the Malaysian Malay and Chinese ethnic groups using the Primer Express software. "Blind tests" using DNA samples from healthy individuals and β-thalassemia patients with different genotypes were performed to determine the specificity and sensitivity of this newly designed assay. Our results showed 100% sensitivity and specificity for this novel assay. In conclusion, the TaqMan genotyping assay is a straightforward assay that allows detection of β-globin gene mutations in less than 40 min. The simplicity and reproducibility of the TaqMan genotyping assay permit its use in laboratories as a rapid and cost-effective diagnostic tool for confirmation of common β-thalassemia mutations in Malaysia. Fundacao de Pesquisas Cientificas de Ribeirao Preto 2013-07-15 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/29670/1/Specific%20and%20straightforward%20molecular%20investigation%20of%20%CE%B2.pdf Kho, Siew Leng and Chua, Kek Heng and George, Elizabeth and Tan, Mary Anne Jin Ai (2013) Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays. Genetics and Molecular Research, 12 (3). pp. 2409-2415. ISSN 1676-5680 http://www.geneticsmr.com/articles/2116 10.4238/2013.February.28.4
spellingShingle Kho, Siew Leng
Chua, Kek Heng
George, Elizabeth
Tan, Mary Anne Jin Ai
Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays
title Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays
title_full Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays
title_fullStr Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays
title_full_unstemmed Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays
title_short Specific and straightforward molecular investigation of β-thalassemia mutations in the Malaysian Malays and Chinese using direct TaqMan genotyping assays
title_sort specific and straightforward molecular investigation of β-thalassemia mutations in the malaysian malays and chinese using direct taqman genotyping assays
url http://psasir.upm.edu.my/id/eprint/29670/
http://psasir.upm.edu.my/id/eprint/29670/
http://psasir.upm.edu.my/id/eprint/29670/
http://psasir.upm.edu.my/id/eprint/29670/1/Specific%20and%20straightforward%20molecular%20investigation%20of%20%CE%B2.pdf