Molecular genetic study on G6PD deficiency in Philippines

本文建立了一种用滤纸点样法测定全血中葡萄糖6磷酸脱氢酶（G6PD）活性筛查G6PD缺乏症的方法。在居住在大马尼拉地区的人群中进行G6 PD缺乏症筛查。约30，000名新生儿进行了G6PD缺乏症筛查，发现3.6%的新生儿患有G6PD缺乏症。虽然超过300种不同的G6PD变异体已被区分的生化参数的基础上提出了巨大的遗传异质性，只有一小部分的这种异质性已被证实在DNA.To促进澄清的分子发病机制的G6PD缺陷，我们已经发明了一种新的突变分析系统命名为MPTP（多重PCR与串联引物）。通过MPTP，可以在两天内通过PCR检测到G6PD基因上的单个核苷酸变化。我们应用MPTP检测了12种不同的突变，并在大多数病例中发现了突变，部分病例MPTP未发现突变，并对G6PD基因编码区进行了全长测序。结果发现了一个新的T198A突变。

A method to screen G6PD deficiency was established by measuring enzyme activity in whole blood spotted on a filter paper. Screening of G6PD deficiency was conducted in peoples living in metro Manila area. Around 30,000 neonates were screened for G6PD deficiency and 3.6% of neonates were found to be G6PD deficiency. Although more than 300 different G6PD variants have been distinguished on the basis of biochemical parameters suggesting a vast genetic heterogeneity, only a small portion of this heterogeneity has been confirmed at the DNA.To facilitate clarification of molecular pathogenesis of G6PD deficiency, we have invented a new mutation analysis system named MPTP (Multiplex PCR with tandem primers). By MPTP a single nucleotide change on the G6PD gene can be detected by doing PCR within two days. We applied MPTP to detect mutations ofl2 different mutations and identified mutations in most of cases.Some cases had no mutation identified by MPTP and the full length of the coding region of the G6PD gene was sequenced. As a result, a novel mutation of T198A was identified.