目的 了解新疆维吾尔自治区（新疆）鼠疫耶尔森菌（鼠疫菌）规律聚集的间隔短回文重复（CRISPR）位点的遗传多态性，对菌株进行基因分型。方法 利用PCR技术对分离自新疆的58株鼠疫菌的基因组DNA进行扩增，测定扩增产物的核酸序列，分析各菌株CRISPR位点的间区序列种类和排列情况，应用Bionumerics软件制作聚类图，确定菌株基因型。结果 共鉴定42个间区序列和23种间区序列阵列，58株鼠疫菌分为16个基因型，即基因3型、8型、11型和16型，主要分布在新疆乌恰县、和田地区、尼勒克和玛纳斯县；基因7型的鼠疫菌全部分离自新疆准噶尔盆地大沙鼠及其体表寄生蚤；其他型别的鼠疫菌则主要分布在新疆北天山和准噶尔盆地。结论 新疆地区的鼠疫菌CRISPR位点遗传多态性较高，不同基因型的鼠疫菌地理分布有明显的区域性特征。
Objective To study the genetic polymorphism of clustered regularly interspaced short palindromic repeat (CRISPR)loci in the Yersinia pestis strains from Xinjiang Uygur Autonomous Region, China and then to identify the genotypes of tested strains. Methods Genomic DNA of 58 Y. pestis strains from Xinjiang was amplified using PCR technique, and the nucleic acid sequences of PCR production were then determined. On each CRISPR locus of every strain, spacers and spacer arrays were analyzed according to the PCR production sequences. Genotypes of tested strains were identified through the dendrogram generated by software. Results A total of 42 spacers and 23 spacer arrays were confirmed and 58 Y. pestis strains were divided into 16 genotypes. Yersinia pestis strains of genotype 3, 8, 11 and 16 distributed intensively in Wuqia county, Hotan prefecture, Nilka county and Manasi county respectively; strains of genotype 7 were all isolated from Rhombomys opimus and parasitic fleas in vitro in Junggar Basin; strains of the other genotypes mainly distributed in North Tianshan and Junggar Basin. Conclusion The genetic polymorphism of CRISPR loci in Xinjiang Y. pestis strains is high, and the geographical distribution of Y. pestis strains of different genotypes has obviously regional characteristics.
[1] 卫生部卫生应急办公室,中国疾病预防控制中心. 鼠疫防控应急手册[M]. 北京:北京大学医学出版社, 2009:1-2.
[2] 张贵军,段天一,邵奎东,等. 全国2015年鼠疫监测结果[J]. 中国地方病防治杂志, 2016, 31增刊:1-7.
[3] 纪树立,张海峻,刘云鹏,等. 我国鼠疫菌分型及其生态学、流行病学意义[J]. 中国地方病学杂志, 1987, 6(5):3-9, 62.
[4] 杨晓艳,魏柏青,靳娟,等. 中国鼠疫耶尔森菌差异区段分型及其地理分布特征[J]. 中华流行病学杂志, 2014, 35(8):943-948.
[5] Pourcel C,Salvignol G,Vergnaud G. CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA,and provide additional tools for evolutionary studies[J]. Microbiology, 2005, 151(3):653-663.
[6] Cui YJ,Li YJ,Gorgé O,et al. Insight into microevolution of Yersinia pestis by clustered regularly interspaced short palindromic repeats[J]. PLoS One, 2008, 3(7):e2652.
[7] 许磊,方喜业,周冬生,等. 中国鼠疫自然疫源地的演化动态及环境生态位的生物学特征[J]. 中国媒介生物学及控制杂志, 2015, 26(3):228-232.
[8] 热娜·吐尔地,布仁明德,戴翔,等. 新疆准噶尔盆地鼠疫菌生化和毒力测定[J]. 中国人兽共患病学报, 2006, 22(11):1086-1087.
