中华卫生杀虫药械

目的 探讨青海省鼠疫自然疫源地鼠疫菌单核苷酸多态性(single nucleotide polymorphism, SNP)的基因型及其地区分布。方法 选取青海省地方病预防控制所保存的1954—2020年在不同地区宿主、媒介体内分离的250株鼠疫菌作为实验菌株，采用传统的苯酚-氯仿混合抽提法提取鼠疫菌DNA。使用Illumina HiSeq 2000平台对250株鼠疫菌进行全基因测序，根据文献中鼠疫菌全基因组SNPs分析方法，基于133株国内外鼠疫菌株及假结核耶尔森菌IP32953,比对2 298个SNP位点，用MEGA 6.0软件对250株青海省鼠疫菌以及32株参考菌株构建系统发育关系。结果 青海省250株鼠疫菌分布于8个市(州),31个县(区)。其中海西州共66株，海北州共70株，海南州共28株，玉树州共62株，黄南州、果洛州、西宁市、海东市共24株。基于2 298个SNPs位点的聚类结果显示，青海省250株鼠疫菌分布在5个亚分支中，分别为0.PE分支6株(2.0%),1.IN分支235株(94.0%),2.ANT分支4株(1.6%),2.MED分支1株(0.4%),3.ANT分支4株(1.6%)。结论 青海省鼠疫自然疫源地鼠疫菌SNP基因型具有高度多样性，地区分布特征明显。

Objective To study the genotype and regional distribution of single nucleotide polymorphism(SNP) of Yersinia pestis in Qinghai Province. Methods A toal of 250 strains of Y. pestis isolated from hosts and vectors in different regions from 1954 to 2020 were selected as experimental strains, the DNA of Y. pestis was extracted by using the traditional phenol-chloroform mixed extraction method. The Illumina HiSeq 2000 platform was used to sequence the whole gene of 250 strains. According to the SNPs analysis method of the whole genome of Y. pestis in the literature, based on 133 domestic and foreign strains of Y. pestis and Y. pseudotuberculosis IP32953, 2 298 SNP loci were compared. MEGA 6.0 software was used to construct phylogenetic relationships for the 250 strains and 32 reference strains of Qinghai Province. Results A total of 250 strains were distributed in 8 cities/states and 31 counties/districts in Qinghai Province. There were 66 strains in Haixi Prefecture, 70 strains in Haibei Prefecture, 28 strains in Hainan Prefecture, 62 strains in Yushu Prefecture, and 24 strains in Huangnan Prefecture, Golog Prefecture, Xining City, and Haidong City. Based on the clustering of 2 298 SNPs sites, the 250 strains of Y. pestis from Qinghai Province were distributed in 5 subclades: 6 strains(2.0%) from 0.PE clade, 235 strains(94.0%) from 1.IN clade, 4 strains(1.6%) from 2.ANT clade, 1 strain(0.4%) from 2.MED clade, and 4 strains of 3.ANT branch(1.6%).Conclusion There is a high diversity of SNP genotypes in plague natural foci of Qinghai Province, and the regional distribution characteristics are obvious.