Abstract：Objective To investigate the genotype of Yersinia pestis isolated from the natural plague focus of Meriones unguiculatus in Inner Mongolian Plateau using clustered regularly interspaced short palindromic repeats (CRISPR) and the difference in the genotype of strains from the same type of epidemic focus and different areas, to establish the CRISPR gene bank of plague strains from gerbil plague foci, and to lay a foundation for epidemiological trace-back and analysis of epidemic situation. Methods Three pairs of CRISPR primers (YPa, YPb, and YPc) were used for PCR amplification and sequencing of the DNA of experimental strains, and the CRISPR sequence obtained was compared with the latest CRISPR Dictionary reported in literature to obtain CRISPR spacer array and identify genotype. Bionumerics 7.6 software was used to plot clustering charts and analyze the phylogenetic relationship. Results A total of 9 spacers were found in 33 strains of Y. pestis, i.e., 4 types of YPa (a1, a2, a3, and a56), 2 types of YPb (b1 and b2), and 3 types of YPc (c1, c2, and c3). The comparative analysis showed that all strains were classified as 1 CRISPR gene cluster (Cb2) with two genotypes. The strains isolated from Etuoke banner and Hanggin Rear banner of Inner Mongolia were identified as a new genotype, which was named as genotype 2' (a1-a2-a3-a56, b1-b2, c1-c2-c3), and the strains from Urad Front banner of Inner Mongolia, Kangbao county of Hebei province, and Yinchuan city of Ningxia Hui autonomous region were identified as genotype 1 (a1-a2-a3, b1-b2, c1-c2-c3). ConclusionY. pestis isolated from the plague foci of M. unguiculatus in Inner Mongolian is genetically stable as a whole and is classified as one gene cluster, but a certain degree of microevolution is observed, which reflects that strains from different areas have different genotypes.
王海峰, 陈永明, 周松, 牛艳芬, 杨晓燕, 张懿晖, 刘广, 杜国义, 刘合智, 史献明. 成簇的规律间隔短回文重复序列技术在鼠疫菌基因分型中的应用[J]. 中国媒介生物学及控制杂志, 2021, 32(1): 30-33.
WANG Hai-feng, CHEN Yong-ming, ZHOU Song, NIU Yan-fen, YANG Xiao-yan, ZHANG Yi-hui, LIU Guang, DU Guo-yi, LIU He-zhi, SHI Xian-ming. Application of clustered regularly interspaced short palindromic repeats in genotyping of Yersinia pestis. Chines Journal of Vector Biology and Control, 2021, 32(1): 30-33.
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2021, Vol. 32 
