Analysis of common mutations in the galactose-1-phosphate uridyl transferase gene: new assays to increase the sensitivity and specificity of newborn screening for galactosemia.
Analysis of common mutations in the galactose-1-phosphate uridyl transferase gene: new assays to increase the sensitivity and specificity of newborn screening for galactosemia.
复制标题
1-磷酸半乳糖尿苷基转移酶基因常见突变分析：提高新生儿半乳糖血症筛查敏感性和特异性的新测定方法。
DOI：
10.1016/s1525-1578(10)60450-3
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Naylor,EdwinW
中科院分区：
文献类型：
--
作者：
Dobrowolski,StevenF;Banas,RichardA;Suzow,JosephG;Berkley,Michelle;Naylor,EdwinW
经典半乳糖血症是由半乳糖-1-磷酸尿苷转移酶（GALT）基因突变引起的遗传性疾病。新生儿半乳糖血症的前瞻性筛查是常规的，并利用普遍收集的新生儿干血样在滤纸上。筛选半乳糖血症是通过分析总半乳糖（半乳糖和半乳糖-1-磷酸）和/或测定GALT酶的活性来实现的。虽然这种方法是有效的，但环境因素和高频率的Duarte D2突变（N314D）确实会导致假阳性结果。利用从原始新生儿干血标本中提取的DNA和Light Cycler技术，描述了能够检测四种最常见的经典半乳糖血症等位基因（Q188R, S135L， K285N和L195P）和N314D Duarte变异突变的五种检测方法。这五种检测方法在常用条件下同时进行。包括DNA制备，建立，扩增和分析，所有五个位点的基因型数据在不到2小时内获得。检测结果易于解释，适用于高通量新生儿筛查。突变分析有助于减少假阳性结果，区分D/G混合杂合子与经典半乳糖血症，并清楚地确定经典半乳糖血症患者的高百分比。
Classical galactosemia is a genetic disease caused by mutations in the galactose-1-phosphate uridyl transferase (GALT) gene. Prospective newborn screening for galactosemia is routine and utilizes the universally collected newborn dried blood specimen on filter paper. Screening for galactosemia is achieved through analysis of total galactose (galactose and galactose-1-phosphate) and/or determining the activity of the GALT enzyme. While this approach is effective, en vironmental factors and the high frequency of the Duarte D2 mutation (N314D) does lead to false positive results. Using DNA derived from the original newborn dried blood specimen and Light Cycler technology a panel of five assays able to detect the four most frequently encountered classical galactosemia alleles (Q188R, S135L, K285N, and L195P) and the N314D Duarte variant mutation are described. The five assays are performed simultaneously using common conditions. Including DNA preparation, set-up, amplification, and analysis the genotype data for all five loci is obtained in less than 2 hours. The assays are easily interpreted and amenable to high-throughput newborn screening. Mutational analysis is useful to reduce false positive results, differentiate D/G mixed heterozygotes from classical galactosemia, and to clearly identify a very high percentage of those affected by classical galactosemia.