云南省输入性及本地感染间日疟原虫裂殖子表面蛋白1基因5区序列的多态性分析

发布时间：2018-02-16 18:17

本文关键词： 间日疟原虫裂殖子表面蛋白多态性　出处：《中国寄生虫学与寄生虫病杂志》2017年01期 　论文类型：期刊论文

【摘要】：目的分析云南省输入性及本地感染间日疟病例的疟原虫裂殖子表面蛋白1(Plasmodium vivax merozoite surface protein-1,Pv MSP-1)基因5区序列及其等位基因的多态性。方法 2012年8月-2015年9月,收集云南省输入性及本地感染间日疟病例的滤纸血样和流行病学史等相关信息。提取疟原虫DNA,PCR扩增Pv MSP-1基因5区并测序,与M75674、AAN86237、M60807、ABJ53045、AAN86238、BAA18977等参比序列进行比对,用Mega 5.04、Arlequin3.5.1软件分析Pv MSP-1基因5区的序列多态性,计算序列间保守位点、遗传距离和多样性指数,并根据氨基酸序列间的遗传距离绘制聚类树状图。结果 2012年8月-2015年9月,共收集间日疟病例血样847份。其中,云南省本地感染61份,非洲、缅甸感染的分别为66和720份。847份血样中278份扩增出Pv MSP-1基因5区片段,206份测序成功。Pv MSP-1基因5区片段编码氨基酸长193~222 aa。氨基酸序列比对分析显示,Sal-1、Belem和Recombine基因型分别占59.2%(122/206)、23.3%(48/206)和17.5%(36/206),缅甸、非洲感染和云南本地感染血样中Sal-1基因型分别占58.8%(104/177)、11/15和7/14。Sal-1、Belem、Recombine基因型分别有51、9和6个亚型。这66个亚型的Shannon-wiener指数(H’)和期望杂合度(He)分别为0.955和0.567。206条DNA序列的同源位点为665 bp、保守位点为11.3%(75/665)、变异位点为88.7%(590/665),不同感染来源地的分离株与不同的氨基酸参比序列的遗传距离均小于0.4。聚类分析显示,206条Pv MSP-1基因5区氨基酸序列按基因型聚类成2个大类,有3个次级分支,其中,Recombine与Belem基因型的相似度为91%~92%,高于与Sal-1基因型的82%~83%。结论云南省输入性及本地感染的间日疟原虫分离株Pv MSP-1基因5区存在多种亚型,在Sal-1、Belem、Recombine等3种基因型中,Sal-1型占优势。
[Abstract]:Objective to analyze the sequence and allele polymorphism of Plasmodium vivax merozoite surface protein-1 PvMSP-1) gene in Plasmodium vivax merozoite surface protein-1PvMSP-1 gene of Plasmodium vivax and Plasmodium merozoite surface protein-1in imported and locally infected Plasmodium vivax malaria cases from August 2012 to September 2015 in Yunnan Province. The paper blood samples and epidemiological history of imported and locally infected Plasmodium vivax malaria cases in Yunnan Province were collected. The Pv MSP-1 gene region 5 was amplified by PCR from Plasmodium falciparum DNA and sequenced, and compared with the reference sequence of A75674 AAN86237 M60807 ABJ53045 AAN86238BAA18977. Mega 5.04 Arlequin3.5.1 software was used to analyze the polymorphism of PV MSP-1 gene region 5, to calculate the conserved loci, genetic distance and diversity index between sequences, and to draw a cluster tree map according to the genetic distance between amino acid sequences. A total of 847 blood samples were collected from vivax malaria cases. Two hundred and eighty-six out of 66 and 720 blood samples from Myanmar were infected with PV MSP-1 gene. Two hundred and six samples were sequenced successfully. The amino acid length of the 5 region fragment of the PV MSP-1 gene was 1993 ~ (aaa). The analysis of amino acid sequence comparison showed that Sal-1Bel and Recombine genotypes were detected by comparison of amino acid sequences. Don't take 59.2% 122.2 / 206 / 23.3t / 206) and 17.5a / 36R / 206b, Burma, In the blood samples of African infection and Yunnan native infection, the proportion of Sal-1 genotypes in 104 / 177G / 11 / 15 and 7 / 14 / Sal-1Bel / Recombine genotypes were 51 / 9 and 6, respectively. The Shannon-wiener index and expected heterozygosity of the 66 subtypes were 0.955 and 0.567.206 DNA homologous sites, respectively. 665 BP, the conserved site was 11.3% / 665N, the mutation site was 88.70.90 / 665N, and the genetic distance between the isolated strains from different infection sources and different amino acid reference sequences was less than 0.4. Cluster analysis showed that the amino acid sequences of the 5 region of Pv MSP-1 gene were clustered into two groups according to genotype. There were three secondary branches, among which the similarity between Recombine and Belem genotype was 91 / 92, which was higher than that with Sal-1 genotype. Conclusion there are many subtypes in Pv MSP-1 gene region 5 of P. vivax isolates isolated from Yunnan Province. The Sal-1 genotype was dominant in three genotypes, such as Sal-1 Belemer Recombine and so on.
【作者单位】：云南省寄生虫病防治所云南省疟疾研究中心云南省虫媒传染病防控研究重点实验室;大理大学基础医学院;
【基金】：国家自然科学基金(No.81220108019,81660559) 上海市2015年度科技创新行动计划(No.15495800400)~~
【分类号】：R382.31

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