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The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations

Introduction: In Malaysia, β-thalassaemia is a common inherited blood disorder in haemoglobin synthesis with a carrier rate of 4.5%. Currently, PCR-incorporating techniques such as amplification refractory mutation system (ARMS) or reverse dot blot hybridization (RDBH) are used in β-thalassaemia mut...

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Egile Nagusiak: Teh, L. K., Lee, T. Y., Tan, Mary Anne Jin Ai, Lai, Mei I., George, E.
Formatua: Artikulua
Hizkuntza:English
Argitaratua: Wiley-Blackwell Publishing 2015
Sarrera elektronikoa:http://psasir.upm.edu.my/id/eprint/46713/1/The%20use%20of%20Taqman%20genotyping%20assays%20for%20rapid%20confirmation%20of%20B-thalassaemia%20mutations%20in%20the%20Malays%3B%20accurate%20diagnosis%20with%20low%20DNA%20concentrations_New.pdf
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spelling oai:psasir.upm.edu.my:46713 http://psasir.upm.edu.my/id/eprint/46713/ The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations Teh, L. K. Lee, T. Y. Tan, Mary Anne Jin Ai Lai, Mei I. George, E. Introduction: In Malaysia, β-thalassaemia is a common inherited blood disorder in haemoglobin synthesis with a carrier rate of 4.5%. Currently, PCR-incorporating techniques such as amplification refractory mutation system (ARMS) or reverse dot blot hybridization (RDBH) are used in β-thalassaemia mutation detection. ARMS allows single-mutation identification using two reactions, one for wild type and another for mutant alleles. RDBH 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 DNA concentrations. Methods: Genotype identification of common β-thalassaemia mutations in Malays was carried out using Taqman genotyping assays. Results: Results show that the Taqman assays allow mutation detection with DNA 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. Conclusion: The developed sensitive Taqman assays allow molecular characterization of β-thalassaemia mutations in samples with low DNA concentrations. The Taqman genotyping assays have potential as a diagnostic tool for foetal blood, chorionic villi or pre-implantation genetic diagnosis where DNA is limited and precious. Wiley-Blackwell Publishing 2015-02 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/46713/1/The%20use%20of%20Taqman%20genotyping%20assays%20for%20rapid%20confirmation%20of%20B-thalassaemia%20mutations%20in%20the%20Malays%3B%20accurate%20diagnosis%20with%20low%20DNA%20concentrations_New.pdf Teh, L. K. and Lee, T. Y. and Tan, Mary Anne Jin Ai and Lai, Mei I. and George, E. (2015) The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations. International Journal of Laboratory Hematology, 37 (1). pp. 79-89. ISSN 1751-5521; ESSN: 1751-553X https://onlinelibrary.wiley.com/doi/10.1111/ijlh.12240 10.1111/ijlh.12240
institution UPM IR
collection UPM IR
language English
description Introduction: In Malaysia, β-thalassaemia is a common inherited blood disorder in haemoglobin synthesis with a carrier rate of 4.5%. Currently, PCR-incorporating techniques such as amplification refractory mutation system (ARMS) or reverse dot blot hybridization (RDBH) are used in β-thalassaemia mutation detection. ARMS allows single-mutation identification using two reactions, one for wild type and another for mutant alleles. RDBH 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 DNA concentrations. Methods: Genotype identification of common β-thalassaemia mutations in Malays was carried out using Taqman genotyping assays. Results: Results show that the Taqman assays allow mutation detection with DNA 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. Conclusion: The developed sensitive Taqman assays allow molecular characterization of β-thalassaemia mutations in samples with low DNA concentrations. The Taqman genotyping assays have potential as a diagnostic tool for foetal blood, chorionic villi or pre-implantation genetic diagnosis where DNA is limited and precious.
format Article
author Teh, L. K.
Lee, T. Y.
Tan, Mary Anne Jin Ai
Lai, Mei I.
George, E.
spellingShingle Teh, L. K.
Lee, T. Y.
Tan, Mary Anne Jin Ai
Lai, Mei I.
George, E.
The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations
author_facet Teh, L. K.
Lee, T. Y.
Tan, Mary Anne Jin Ai
Lai, Mei I.
George, E.
author_sort Teh, L. K.
title The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations
title_short The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations
title_full The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations
title_fullStr The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations
title_full_unstemmed The use of Taqman genotyping assays for rapid confirmation of B-thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations
title_sort use of taqman genotyping assays for rapid confirmation of b-thalassaemia mutations in the malays: accurate diagnosis with low dna concentrations
publisher Wiley-Blackwell Publishing
publishDate 2015
url http://psasir.upm.edu.my/id/eprint/46713/1/The%20use%20of%20Taqman%20genotyping%20assays%20for%20rapid%20confirmation%20of%20B-thalassaemia%20mutations%20in%20the%20Malays%3B%20accurate%20diagnosis%20with%20low%20DNA%20concentrations_New.pdf
_version_ 1819296745402662912
score 13.4562235

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