The use of high resolution melting analysis to detect Fabry mutations in heterozygous females via dry bloodspots

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• 作者：Chang-Long Tai^a ; ^b ; ¹ ; Mei-Ying Liu^a ; ¹ ; Hsiao-Chi Yu^a ; Chiang-Chuan Chiang^c ; Hung Chiang^d ; Jeng-Hung Suen^d ; Shu-Min Kao^c ; Yu-Hsiu Huang^a ; Tina Jui-Ting Wu^a ; Chia-Feng Yang^a ; Fang-Chih Tsai^a ; Ching-Yuang Lin^e ; Jan-Gowth Chang^f ; Hong-Duo Chen^b ; ^{hongduochen@hotmail.com} ; Dau-Ming Niu^a ; ^g ; ^{dmniu1111@yahoo.com.tw}
• 关键词：Fabry disease ; GLA ; High-resolution melting analysis ; Heterozygous female
• 刊名：Clinica Chimica Acta
• 出版年：2012
• 出版时间：18 February 2012
• 年：2012
• 卷：413
• 期：3-4
• 页码：422-427
• 全文大小：1142 K

文摘

Background

As an X-linked genetic disorder, Fabry disease was first thought to affect males only, and females were generally considered to be asymptomatic carriers. However, recent research suggests that female carriers of Fabry disease may still develop vital organ damage causing severe morbidity and mortality. In the previous newborn screening, from 299,007 newborns, we identified a total of 20 different Fabry mutations and 121 newborns with Fabry mutations. However, we found that most female carriers are not detected by enzyme assays.

Methods

A streamlined method for high resolution melting (HRM) analysis was designed to screen for GLA gene mutations using a same PCR and melting program. Primer sets were designed to cover the 7 exons and the Chinese common intronic mutation, IVS4+919G>A of GLA gene.

Results

The HRM analysis was successful in identifying heterozygous and hemizygous patients with the 20 surveyed mutations. We were also successful in using this method to test dry blood spots of newborns afflicted with Fabry mutations without having to determine DNA concentration before PCR amplification.

Conclusion

The results of this study show that HRM could be a reliable and sensitive method for use in the rapid screening of females for GLA mutations.