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PLoS One. 2014 Aug 4;9(8):e102243. doi: 10.1371/journal.pone.0102243. eCollection 2014.

High resolution melting analysis: a rapid screening and typing tool for common β-thalassemia mutation in Chinese population.

Author information

  • 1Central Laboratory, Chaozhou Central Hospital Affiliated to Southern Medical University, Chaozhou, Guangdong Province, People's Republic of China; Medical Laboratory, First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong Province, People's Republic of China.
  • 2Department of Biology, Hanshan Normal University, Chaozhou, Guangdong Province, People's Republic of China.
  • 3Central Laboratory, Chaozhou Central Hospital Affiliated to Southern Medical University, Chaozhou, Guangdong Province, People's Republic of China.
  • 4Medical Laboratory, First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong Province, People's Republic of China.
  • 5Department of Medical Laboratory, Hospital of Youjiang Medical University for Nationalities, Baise, Guangxi Province, People's Republic of China.
  • 6Medical Laboratory, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi Province, People's Republic of China.
  • 7Laboratory Medical Center, People's Hospital of Yunan Province, Kunming, Yunnan Province, People's Republic of China.
  • 8Department of Medical Laboratory, Chengdu Women's & Children's Central Hospital, Chengdu, Sichuan Province, People's Republic of China.
  • 9Central Laboratory, Chaozhou Central Hospital Affiliated to Southern Medical University, Chaozhou, Guangdong Province, People's Republic of China; Laboratory Medical Center, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong Province, People's Republic of China.
  • 10Laboratory Medical Center, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong Province, People's Republic of China.
  • 11Medical Laboratory, Meixian People's Hospital, Meizhou, Guangdong Province, People's Republic of China.

Abstract

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.

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