High Resolution Melting Analysis: A Rapid Screening and Typing Tool for Common [beta]-Thalassemia Mutation in Chinese Population

[beta]-thalassemia is a common inherited disorder worldwide including southern China, and at least 45 distinct [beta]-thalassemia mutations have been identified in China. High-resolution melting (HRM) assay was recently introduced as a rapid, inexpensive and effective method for genotyping. However,...

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Veröffentlicht in:PloS one 2014-08, Vol.9 (8)
Hauptverfasser: Lin, Min, Jiao, Ji-Wei, Zhan, Xiu-Hui, Zhan, Xiao-Fen, Pan, Mei-Chen, Wang, Jun-Li, Wang, Chun-Fang, Zhong, Tian-Yu, Zhang, Qin, Yu, Xia, Wu, Jiao-Ren, Yang, Hui-Tian, Lin, Fen, Tong, Xin, Yang, Hui, Zha, Guang-Cai, Wang, Qian, Zheng, Lei, Wen, Ying-Fang, Yang, Li-Ye
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Sprache:eng
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Zusammenfassung:[beta]-thalassemia is a common inherited disorder worldwide including southern China, and at least 45 distinct [beta]-thalassemia mutations have been identified in China. High-resolution melting (HRM) assay was recently introduced as a rapid, inexpensive and effective method for genotyping. However, there was no systemic study on the diagnostic capability of HRM to identify [beta]-thalassemia. Here, we used an improved HRM method to screen and type 12 common [beta]-thalassemia mutations in Chinese, and the rapidity and reliability of this method was investigated. The whole PCR and HRM procedure could be completed in 40 min. The heterozygous mutations and 4 kinds of homozygous mutations could be readily differentiated from the melting curve except c.-78A>G heterozygote and c.-79A>G heterozygote. The diagnostic reliability of this HRM assay was evaluated on 756 pre-typed genomic DNA samples and 50 cases of blood spots on filter paper, which were collected from seven high prevalent provinces in southern China. If c.-78A>G heterozygote and c.-79A>G heterozygote were classified into the same group (c.-78&79 A>G heterozygote), the HRM method was in complete concordance with the reference method (reverse dot blot/DNA-sequencing). In a conclusion, the HRM method appears to be an accurate and sensitive method for the rapid screening and identification of [beta]-thalassemia mutations. In the future, we suggest this technology to be used in neonatal blood spot screening program. It could enlarge the coverage of [beta]-thalassemia screening program in China. At the same time, its value should be confirmed in prospectively clinical and epidemiological studies.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0102243