中缅边境疟疾患者G6PD基因突变型及恶性疟原虫基因分型研究

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common X-linked incomplete dominance inherited disease, G6PD deficient individuals cause hemolytic anemia in clinical manifestations, which is induced by anti-malarial drugs and infection. G6PD gene mutations are the molecular basis of G6PD deficiency, and G6PD gene mutations are geographical and ethnical different. G6PD deficiency has higher incidence rate in malaria-endemic areas due to the fact that it has selective advantage to falciparum malaria. We aim to detect falciparum malaria by microscopy, and screen glucose-6-phosphate dehydrogenase deficiency among pernicious malaria patients by using fluorescent spot method, in order to determine the incidence of G6PD deficiency of pernicious malaria patients, at malaria endemic areas in which near China-Myanmar border. Also we will use the methods of amplify refractory mutation system (ARMS), and denaturing high performance liquid chromatography (DHPLC), to identify chinese known G6PD gene mutations and analyse G6PD gene mutations distribution in that region. At the same time, we want to genotyping Plasmodium falciparum MSP1, MSP2 and GLURP, and obtaining data of their genotype and population characteristics, to explore the relation between G6PD gene mutations distribution to Plasmodium falciparum genotype and population in that region.

葡萄糖6磷酸脱氢酶缺乏症是常见的X连锁不完全显性遗传病，临床上抗疟药物、感染等诱因导致G6PD缺乏症个体发生溶血性贫血等疾病。G6PD基因突变是G6PD缺乏症发生的分子基础，G6PD基因突变型有地域和人种差异。G6PD基因突变后对恶性疟疾有选择优势，因此疟疾流行区G6PD缺乏症发病率偏高。本研究在云南中缅边境疟疾流行区应用镜检恶性疟原虫确定恶性疟疾患者，在恶性疟疾患者中应用荧光斑点法筛查G6PD缺乏症，确定恶性疟疾患者中G6PD缺乏症发生率。应用等位基因特异性扩增（ARMS）结合变性高效液相色谱（DHPLC）方法检测患者中国人已知G6PD基因突变型，分析该地区恶性疟疾患者中G6PD基因突变型的分布特点。同时进行患者恶性疟原虫MSP1、MSP2 和GLURP基因分型，确定患者感染恶性疟原虫的种群特点，探讨该地区恶性疟疾患者G6PD基因突变型分布及与恶性疟原虫基因型、种群的关系。

中缅边境是我国主要的疟疾流行区，由于疟疾的选择压力，导致当地G6PD缺乏症高发。该项目荧光斑点法对中缅边境群体G6PD缺乏症发生频率进行调查，确定中缅边境G6PD缺乏症发生率为29.55%，疟疾和疟疾史病人中G6PD基因缺乏症发生率为10.5% 。项目确定了中缅边境疟疾流行区常见的G6PD基因突变型分布，单体型1311C>T/IVS1193 T>C和487G>A/1311C>T/IVS1193T>C及单碱基突变 G6PD Mahidol 487G>A是中缅边境最常见的三种G6PD突变型。应用巢式PCR完成25例恶性疟疾并G6PD基因突变、48例对照恶性疟原虫MSP1三个位点（K1, MAD20, RO33）、MSP2两个位点（FC27, 3D7/IC1）和GLURP基因分型；完成86例间日疟并发G6PD基因突变、252例间日疟对照疟原虫PvMSP3α基因分型，探讨G6PD基因突变与疟原虫种群的关系。同时，该项目还进行中缅边境G6PD缺乏症与其他血红蛋白病的共遗传研究，为当地高发血液遗传病的诊断和预防提供基础资料。