应用高分辨率熔解分析快速检测两种常见的 G6PD 变异型，即非洲型（A-）和地中海型。
Rapid genotyping of two common G6PD variants, African (A-) and Mediterranean, by high-resolution melting analysis.
机构信息
Unité de Pathologie Moléculaire du Globule Rouge, Fédération de Biochimie et de Biologie Spécialisée, Hôpital Edouard Herriot, Hospices Civils and Université Claude Bernard-Lyon 1, Lyon, France.
出版信息
Clin Biochem. 2010 Jan;43(1-2):193-7. doi: 10.1016/j.clinbiochem.2009.09.012. Epub 2009 Sep 24.
OBJECTIVES
The Mediterranean and A(-) G6PD variants are particularly prevalent in Africa and Southern Europe. Our study was aimed to develop an assay for the rapid genotyping of these two variants by HRM.
METHODS
After PCR reactions corresponding to the G6PD Mediterranean (exon 6), G6PD (A-) (exon 4) and G6PD (A-) (exon 5) mutations, amplicons were submitted to HRM. This protocol was applied to a cohort of 132 patients suffering from sickle cell disease.
RESULTS
Wild, homozygous or hemizygous and heterozygous states were fully discriminated by HRM for all three mutations. HRM results were in total accordance with DNA sequencing for 22 patients of our cohort with a 'A' genotype: presence of the (A-) (exon 5) mutation but absence of the (A-) (exon 4) mutation.
CONCLUSIONS
Our HRM protocols allow a rapid, simple and cost-effective screening of G6PD deficiency in patients originating from the Mediterranean and the African areas.
目的
地中海和 A(-) G6PD 变体在非洲和南欧尤为普遍。本研究旨在通过高分辨率熔解曲线 (HRM) 开发一种快速检测这两种变体的基因分型方法。
方法
在进行对应于 G6PD 地中海（exon 6）、G6PD（A-）（exon 4）和 G6PD（A-）（exon 5）突变的 PCR 反应后，将扩增子提交进行 HRM。该方案应用于 132 名患有镰状细胞病的患者队列。
结果
对于所有三种突变，HRM 完全区分了野生型、纯合子或半合子和杂合子状态。对于我们队列中 22 名具有“A”基因型的患者，HRM 结果与 DNA 测序完全一致：存在（A-）（exon 5）突变，但不存在（A-）（exon 4）突变。
结论
我们的 HRM 方案可快速、简单且具有成本效益地筛查来自地中海和非洲地区的患者的 G6PD 缺乏症。
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