| 苏永强,郭丽民,葛亚俊,席进孝,王宇萌,苗克军,吴斌,徐大琴.甘肃省鼠疫耶尔森菌规律成簇间隔短回文重复序列位点多态性分析及地区分布[J].中华流行病学杂志,2020,41(12):2125-2130 |
| 甘肃省鼠疫耶尔森菌规律成簇间隔短回文重复序列位点多态性分析及地区分布 |
| Analysis on clustered regularly interspaced short palindromic repeats loci polymorphism of Yersinia pestis and its area distribution in Gansu province |
| 收稿日期:2020-01-07 出版日期:2020-12-25 |
| DOI：10.3760/cma.j.cn112338-20200107-00017 |
| 中文关键词: 鼠疫耶尔森菌 规律成簇的间隔短回文重复序列 基因型 |
| 英文关键词: Yersinia pestis Clustered regularly interspaced short palindromic repeats Genotyping |
| 基金项目:国家自然科学基金（81560541）；甘肃省卫生行业计划（GSWSKY-2017-21） |
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| 中文摘要: |
| 目的 研究甘肃省鼠疫耶尔森菌（鼠疫菌）的规律成簇的间隔短回文重复序列（CRISPR）位点多态性及地区分布。方法 选取1962－2014年分离的203株鼠疫菌，培养并提取DNA。采用3对CRISPR引物对菌株DNA进行PCR扩增，对PCR产物进行测序。根据菌株CRISPR位点的间区序列种类和排列情况，确定菌株的基因型（类群），采用BioNumerics 5.10软件进行聚类分析。结果 203株鼠疫菌发现有16种间区序列，包括YPa位点9种、YPb位点4种、YPc位点3种，发现新的间区序列a1'。共发现5个CRISPR基因簇，分别为Cb2、Ca7、Ca7'、CaΔ5'、Ca35'。不同的基因簇呈现区域性特征：Cb2主要分布在会宁县、平川区，Ca7主要分布阿克塞哈萨克族自治县；Ca7'主要分布在夏河县；Ca35'主要分布肃北蒙古族自治县、玉门市；CaΔ5'主要集中在肃南裕固族自治县。结论 CRISPR分子分型方法能够较好地区分甘肃省不同疫源地的菌株，且各基因簇呈现一定的区域性特征。对研究甘肃省鼠疫菌进化规律和人间疫情分子生物学溯源有一定意义。 |
| 英文摘要: |
| Objective To study the clustered regularly interspaced short palindromic repeats (CRISPR) loci polymorphism of Yersinia pestis and its area distribution in Gansu province. Methods A total of 203 strains of Yersinia pestis isolated from 1962 to 2014 were selected for the culture and extraction of DNA. Three pairs of CRISPR primers were used to amplify the strain DNA by PCR, and the PCR products were sequenced. The groups and genotypes of strains were determined according to the spacer and spacer arrangement of CRISPR loci in the strain. Cluster analysis was done by using the software BioNumerics 5.10. Results A total of 16 spacers, including 9 species of YPa loci, 4 species of YPb loci and 3 species of YPc loci, were found in the 203 strains of Yersinia pestis. A new spacer of a1' was found. The 203 strains were divided into 5 CRISPR genotypes and classified into 5 CRISPR clusters (Cb2, Ca7, Ca7', CaΔ5' and Ca35'). Each cluster showed significant area-specific characteristics, Cb2 was mainly distributed in Huining country and Pingchuan district, Ca7 was mainly found in Aksai Kazak autonomous country, Ca7' was mainly found in Xiahe country, Ca35' was mainly found in Subei Mongolia autonomous county and Yumen city and CaΔ5' was mainly distributed in Sunan Yugur autonomous county. Conclusions The strains from different plague foci in Gansu were distinguished by CRISPR, all kinds of clusters showed the obvious area specific characteristics. It is important to study the evolution of Yersinia pestis in Gansu and trace the molecular biology origin of human plague. |
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