不同地理株疟原虫裂殖子顶端膜抗原1ama-1基因多态性分析-polymorphism analysis of plasmodium merozoite apical membrane antigen 1ama - 1 gene in different geographical strains.docxVIP

PAGE PAGE 10 间日疟原虫样本顶端膜抗原 1（AMA-1）Ⅱ区具有较低的核苷酸和单倍型多 态性，不同地理株间差异不大，因此用于了解间日疟原虫的遗传特征意义不大。 PvAMA-1Ⅰ区基因可能存在负向选择，而且较少发生减数重组。 研究结果为进一步了解疟疾流行特征，制定疟疾防制措施以及疟疾疫苗的研 制提供分子基础。 【关键词】 恶性疟原虫，间日疟原虫，顶端膜抗原 1，基因多态性 Study on genetic diversity of plasmodium AMA-1 among different strain Abstract 【Objective】 Study on the genetic diversity of Plasmodium falciparum apical membrane antigen 1 (PfAMA-1) and the genetic diversity of Plasmodium vivax apical membrane antigen 1 (PvAMA-1) to understand the genetic polymorphism and distribution at AMA-1. Explore the reasons of maintaining genetic polymorphism at AMA-1, which should be make a molecular genetics basis for further understanding the epidemiological characteristics of malaria , prevention and control measures of malaria and vaccine development using this antigen. 【Methods】 After patients infected by plasmodium were confirmed by microscopy, filter blood samples were collected from patients who attended. The individual information of malaria cases were registered detailly. DNA was extracted from filter blood samples using Na2HPO4 method, nested PCR was employed for amplification the domainⅡ of plasmodium vivax AMA-1 and domainⅠ of Plasmodium falciparum AMA-1. The PCR products were Electrophoresed on agarose gel. Then PCR products which had the right DNA band were purified and sequenced. The sequenced result was analyzed using biosoftware. 【Results】 Analyses of all 23 PfAMA-1 sequences resulted into total 18 haplotypes, where 8 haplotypes were not found in the GenBank database. we found 32 segregating site and the value of nucleotide diversity was 0.02501±0.00099.The average difference of dN-dS for all 23 PfAMA-1 sequences was 0.031±0.006 and All neutrality tests were not significant. The minimum number of recombination events (Rm) was 10. The LD index R2 evidently declined with increasing nucleotide distance. Analyses of all 17 PvAMA-1 sequences resulted into total 9 haplotypes, where only 1 haplotype was new and not found in the GenBank