Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia
Introduction: Beta-thalassaemia is an autosomal recessive disorder and it is a public health problem in the Malaysian Malays and Chinese. This disorder mainly results from point mutations, small insertion or deletions in the β-globin gene complex. Beta-thalassaemia major patients require life-long...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2014
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| Online Access: | http://psasir.upm.edu.my/id/eprint/34570/1/Molecular%20characterisation%20of%20%CE%B2-globin%20gene%20mutations%20in%20Penang%20and%20Kedah%2C%20Malaysia.pdf |
| _version_ | 1796972531222052864 |
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| author | Kho, Siew Leng Chua, Kek Heng George, Elizabeth Goh, A. S. Tan, Mary Anne Jin Ai |
| author_facet | Kho, Siew Leng Chua, Kek Heng George, Elizabeth Goh, A. S. Tan, Mary Anne Jin Ai |
| author_sort | Kho, Siew Leng |
| collection | UPM |
| description | Introduction:
Beta-thalassaemia is an autosomal recessive disorder and it is a public health problem in the Malaysian Malays and Chinese. This disorder mainly results from point mutations, small insertion or deletions in the β-globin gene complex. Beta-thalassaemia major patients require life-long monthly blood transfusions and iron-chelation therapies to sustain their lives. Mutation characterisation is necessary for affected couples at risk of having a β-thalassaemia major child.
Objective:
1. To develop the TaqMan genotyping platform as a time- and cost-effective approach for characterisation of β-globin gene mutations. 2. To characterise the mutations using the developed assays in transfusion-dependent patients in Penang and Kedah.
Methods:
Ten sets of primers and TaqMan probes were designed to identify the common mutations in Malaysian Malays and Chinese: −28 (A→G), CD17 (A→T), CD19 (A→G), HbE (G→A), IVS1-1 (G→T), IVS1-5 (G→C), CD 41/42 (-CTTT), CD71/72 (+A), IVS2-654 (C→T) and Poly A (AATAAAHAATAGA). Another 7 sets of TaqMan genotyping assays were designed to identify the rare mutations in Malays and Chinese: −29 (A→G), Cap (+1) (A→C), CD8/9 (+G), CD16 (-C), CD27/28 (+C), IVS1-1 (G→A) and CD43 (G→T). The developed assays were used to screen 54 and 62 transfusion-dependent patients in Penang and Kedah respectively.
Results & Discussion:
The developed assays detected 92.9% of mutations in the β-thalassaemia major patients. The remaining mutations were detected by ARMS, gap-PCR and DNA sequencing. The most common mutation in β-thalassaemia major patients in Penang is CD41/42 with a frequency of 20.9%. The most common mutation in β-thalassaemia major patients in Kedah is HbE with a frequency of 30.8%.
Conclusion:
The simplicity and reproducibility of the TaqMan genotyping assays enable rapid and cost-effective analysis of the β-globin gene mutations in Malaysia. |
| first_indexed | 2024-03-06T08:29:24Z |
| format | Article |
| id | upm.eprints-34570 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T08:29:24Z |
| publishDate | 2014 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | upm.eprints-345702016-11-30T02:44:51Z http://psasir.upm.edu.my/id/eprint/34570/ Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia Kho, Siew Leng Chua, Kek Heng George, Elizabeth Goh, A. S. Tan, Mary Anne Jin Ai Introduction: Beta-thalassaemia is an autosomal recessive disorder and it is a public health problem in the Malaysian Malays and Chinese. This disorder mainly results from point mutations, small insertion or deletions in the β-globin gene complex. Beta-thalassaemia major patients require life-long monthly blood transfusions and iron-chelation therapies to sustain their lives. Mutation characterisation is necessary for affected couples at risk of having a β-thalassaemia major child. Objective: 1. To develop the TaqMan genotyping platform as a time- and cost-effective approach for characterisation of β-globin gene mutations. 2. To characterise the mutations using the developed assays in transfusion-dependent patients in Penang and Kedah. Methods: Ten sets of primers and TaqMan probes were designed to identify the common mutations in Malaysian Malays and Chinese: −28 (A→G), CD17 (A→T), CD19 (A→G), HbE (G→A), IVS1-1 (G→T), IVS1-5 (G→C), CD 41/42 (-CTTT), CD71/72 (+A), IVS2-654 (C→T) and Poly A (AATAAAHAATAGA). Another 7 sets of TaqMan genotyping assays were designed to identify the rare mutations in Malays and Chinese: −29 (A→G), Cap (+1) (A→C), CD8/9 (+G), CD16 (-C), CD27/28 (+C), IVS1-1 (G→A) and CD43 (G→T). The developed assays were used to screen 54 and 62 transfusion-dependent patients in Penang and Kedah respectively. Results & Discussion: The developed assays detected 92.9% of mutations in the β-thalassaemia major patients. The remaining mutations were detected by ARMS, gap-PCR and DNA sequencing. The most common mutation in β-thalassaemia major patients in Penang is CD41/42 with a frequency of 20.9%. The most common mutation in β-thalassaemia major patients in Kedah is HbE with a frequency of 30.8%. Conclusion: The simplicity and reproducibility of the TaqMan genotyping assays enable rapid and cost-effective analysis of the β-globin gene mutations in Malaysia. Elsevier 2014-06 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/34570/1/Molecular%20characterisation%20of%20%CE%B2-globin%20gene%20mutations%20in%20Penang%20and%20Kedah%2C%20Malaysia.pdf Kho, Siew Leng and Chua, Kek Heng and George, Elizabeth and Goh, A. S. and Tan, Mary Anne Jin Ai (2014) Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia. Asian Pacific Journal of Tropical Disease, 4 (3). p. 232. ISSN 2222-1808 http://www.sciencedirect.com/science/article/pii/S2222180814605284 10.1016/S2222-1808(14)60528-4 |
| spellingShingle | Kho, Siew Leng Chua, Kek Heng George, Elizabeth Goh, A. S. Tan, Mary Anne Jin Ai Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia |
| title | Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia |
| title_full | Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia |
| title_fullStr | Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia |
| title_full_unstemmed | Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia |
| title_short | Molecular characterisation of β-globin gene mutations in Penang and Kedah, Malaysia |
| title_sort | molecular characterisation of β globin gene mutations in penang and kedah malaysia |
| url | http://psasir.upm.edu.my/id/eprint/34570/1/Molecular%20characterisation%20of%20%CE%B2-globin%20gene%20mutations%20in%20Penang%20and%20Kedah%2C%20Malaysia.pdf |
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