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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 polymorphism of Plasmodium vivax (PP) merozoite apical membrane antigen 1 (AMA-1) gene in different geographical isolates. Methods Three drops of filter paper blood were collected from patients with Plasmodium vivax, and pv DNA was extracted from the filter paper by boiling method. DNA fragment containing PvamA-1Ⅱ (793 bp) was amplified by nested PCR. The amplified product was identified by electrophoresis, purified and sequenced , The use of biological software sequence alignment analysis. Results Seventeen patients were infected with 9 haplotypes and only 1 (V3) did not find a 100% anastomosis in the gene pool. Nine polymorphic sites (s = 9), nuclear The nucleotide diversity (π) was 0.00859 ± 0.00093. The difference between the non-synonymous mutation rate and the synonymous mutation rate (dN-dS) was -0.00230 ± 0.00539, but there was no significant difference in Z test (P> 0.05). Neutral test differences were not statistically significant (P> 0.05). The minimum number of recombination events (Rm) was 2, and R2 showed a decreasing trend with the increase of genetic distance in the entire 411bp sequence. Conclusion The genetic diversity of Plasmodium vivax antigen 1 (AMA-1) Ⅱ region is low.