间日疟原虫顶端膜抗原1基因多态性分析  

Study of the genetic diversity of Plasmodium vivax AMA-1

作　　者：周银发[1] 张山鹰[1] 杨发柱[1] 谢汉国[1] 林耀莹[1] 肖方震[1] 

机构地区：[1]福建省疾病预防控制中心,福建福州350001

出　　处：《中国病原生物学杂志》2015年第9期801-804,共4页Journal of Pathogen Biology

基　　金：福建省自然科学基金项目(No.Z0516066)

摘　　要：目的研究不同地理株间日疟原虫(Pv)裂殖子顶端膜抗原1(AMA-1)基因多态性。方法采集镜检确诊的间日疟患者滤纸血3滴,煮沸法提取滤纸血中pv DNA,套式PCR扩增包含PvAMA-1Ⅱ区(793bp)DNA片段,扩增产物经电泳鉴定后纯化并测序,利用生物软件进行序列比对分析。结果 17例患者感染的间日疟原虫分为9种单倍型,只有1种(V3)在基因库中未发现100%吻合的序列;含9个多态位点(s=9),核苷酸多样度(π)为0.00859±0.00093,非同义突变率与同义突变率差值(dN-dS)为-0.00230±0.00539,但Z检验差异无统计学意义(P>0.05)。中性检验差异均无统计学意义(P>0.05)。重组事件的最低数量(Rm)为2,在整个411bp序列中R2随着核苷酸遗传距离增加呈下降趋势不明显。结论间日疟原虫顶端膜抗原1(AMA-1)Ⅱ区基因序列遗传多样性程度较低。Objective To study the genetic diversity of Plasmodium vivax apical membrane antigen 1 in different re- gions. Methods Blood samples were collected froml7 patients infected with Plasmodium vivax, the presence of the pathogen was verified using microscopy, and then the samples were filtered. DNA was extracted by boiling filtered blood samples, and nested PCR was used to amplify the gene fragment （793 bp） containing domain II （411 bp） of Plasmodium vivax AMA-1. The PCR products were electrophoresed on 2% agarose gel. PCR products producing the correct DNA band were purified and sequenced. The sequencing results were analyzed using biosoftware （MEGA4. 10 and DNaSP5. 10）. Results Analyses of all 17 PvAMA-1 sequences resulted in a total of 9 haplotypes. Of these, only V3 lacked a haplotype 100~ similar to haplotypes in the GenBank database. Seven polymorphic loci were identified in the 137 amino acid residues encoded by the domain II gene fragment （411 bp）. Nine segregating sites were identified and nucleotide di- versity was 0. 00859~0. 00093. The average difference in dN-dS for all 17 PvAMA-1 sequences was-0. 00230-4-0. 00539 but Z test results were not significant （P〉0.05）. None of the results of neutrality tests （Tajima＇s test and Fu and Li＇s test） were significant （P〉0.05）. The minimum number of recombination events （Rm） was 2. For the entire 411-bp sequence, the LD index R2 did not markedly decrease with a greater nucleotide distance. Conclusion Domain II of PvA- MA-1 displays little genetic diversity and haplotype diversity.

关 键 词：疟原虫 间日 顶端膜抗原1 基因多态性 

分 类 号：R382.31[医药卫生—医学寄生虫学]