Identification of fibrillin-1 gene mutations in Marfan syndrome by high-resolution melting analysis

Chia Cheng Hung, Shin Yu Lin, Chien Nan Lee, Hui Yu Cheng, Chiou Ya Lin, Chien Hui Chang, Hsin Hui Chiu, Chih Chieh Yu, Shuan Pei Lin, Wen Fang Cheng, Hong Nerng Ho, Dau Ming Niu, Yi Ning Su

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)


Marfan syndrome has been associated with approximately 562 mutations in the fibrillin-1 (FBN1) gene. Mutation scanning of the FBN1 gene with DNA direct sequencing is time-consuming and expensive because of its large size. This study analyzed the diagnostic value of high-resolution melting analysis as an alternative method for scanning of the FBN1 gene. A total of 75 polymerase chain reaction (PCR) amplicons (179-301 bp, average 256 bp) that covered the complete coding regions and splicing sites were evaluated on the 96-well LightCycler system. Melting curves were analyzed as fluorescence derivative plots (-dF/dT vs. temperature). To determine the sensitivity of this method, a total of 82 samples from patients with Marfan syndrome and 50 unaffected individuals were analyzed. All mutations reported in this study had been confirmed previously by direct sequencing analysis. Melting analysis identified 48 heterozygous variants. The variant c.3093 G>T (exon 25) was incorrectly identified by melting curve analysis. The sensitivity of the technique in this sample was 98.78% (81/82). This study demonstrated that high-resolution melting analysis is a reliable gene scanning method with greater speed than DNA sequencing. Our results support the use of this technology as an alternative method for the diagnosis of Marfan syndrome as well as its suitability for high-throughput mutation scanning of other large genes.

Original languageEnglish
Pages (from-to)102-106
Number of pages5
JournalAnalytical Biochemistry
Issue number2
Publication statusPublished - Jun 15 2009
Externally publishedYes


  • FBN1 gene
  • High-resolution melting (HRM)
  • Marfan syndrome

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry
  • Cell Biology


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