High resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population.
β-thalassemia is a common inherited disorder worldwide including southern China, and at least 45 distinct β-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...
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2014
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oai:doaj.org-article:557e147815ce4f868f6269ed2c323dd52021-11-25T06:06:08ZHigh resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population.1932-620310.1371/journal.pone.0102243https://doaj.org/article/557e147815ce4f868f6269ed2c323dd52014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25089872/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203β-thalassemia is a common inherited disorder worldwide including southern China, and at least 45 distinct β-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 β-thalassemia. Here, we used an improved HRM method to screen and type 12 common β-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 β-thalassemia mutations. In the future, we suggest this technology to be used in neonatal blood spot screening program. It could enlarge the coverage of β-thalassemia screening program in China. At the same time, its value should be confirmed in prospectively clinical and epidemiological studies.Min LinJi-Wei JiaoXiu-Hui ZhanXiao-Fen ZhanMei-Chen PanJun-Li WangChun-Fang WangTian-Yu ZhongQin ZhangXia YuJiao-Ren WuHui-Tian YangFen LinXin TongHui YangGuang-Cai ZhaQian WangLei ZhengYing-Fang WenLi-Ye YangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 8, p e102243 (2014) |
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Medicine R Science Q Min Lin Ji-Wei Jiao Xiu-Hui Zhan Xiao-Fen Zhan Mei-Chen Pan Jun-Li Wang Chun-Fang Wang Tian-Yu Zhong Qin Zhang Xia Yu Jiao-Ren Wu Hui-Tian Yang Fen Lin Xin Tong Hui Yang Guang-Cai Zha Qian Wang Lei Zheng Ying-Fang Wen Li-Ye Yang High resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population. |
| description |
β-thalassemia is a common inherited disorder worldwide including southern China, and at least 45 distinct β-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 β-thalassemia. Here, we used an improved HRM method to screen and type 12 common β-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 β-thalassemia mutations. In the future, we suggest this technology to be used in neonatal blood spot screening program. It could enlarge the coverage of β-thalassemia screening program in China. At the same time, its value should be confirmed in prospectively clinical and epidemiological studies. |
| format |
article |
| author |
Min Lin Ji-Wei Jiao Xiu-Hui Zhan Xiao-Fen Zhan Mei-Chen Pan Jun-Li Wang Chun-Fang Wang Tian-Yu Zhong Qin Zhang Xia Yu Jiao-Ren Wu Hui-Tian Yang Fen Lin Xin Tong Hui Yang Guang-Cai Zha Qian Wang Lei Zheng Ying-Fang Wen Li-Ye Yang |
| author_facet |
Min Lin Ji-Wei Jiao Xiu-Hui Zhan Xiao-Fen Zhan Mei-Chen Pan Jun-Li Wang Chun-Fang Wang Tian-Yu Zhong Qin Zhang Xia Yu Jiao-Ren Wu Hui-Tian Yang Fen Lin Xin Tong Hui Yang Guang-Cai Zha Qian Wang Lei Zheng Ying-Fang Wen Li-Ye Yang |
| author_sort |
Min Lin |
| title |
High resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population. |
| title_short |
High resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population. |
| title_full |
High resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population. |
| title_fullStr |
High resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population. |
| title_full_unstemmed |
High resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population. |
| title_sort |
high resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in chinese population. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2014 |
| url |
https://doaj.org/article/557e147815ce4f868f6269ed2c323dd5 |
| work_keys_str_mv |
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