dndC基因在脉冲场凝胶电泳基因组降解中的作用及降解处理方法  

Role of dndC gene in DNA degeneration in pulsed-field gel electrophoresis and treatment method

作　　者：濮吉[1] 周娟[1] 赫自强 熊衍文[1] 金东[1] Pu Ji;Zhou Juan;He Ziqiang;Xiong Yanwen;Jin Dong(State Key Laboratory for Infectious Disease Prevention and Control,National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室,北京102206

出　　处：《疾病监测》2022年第6期796-801,共6页Disease Surveillance

基　　金：海关总署项目(No.2019HK125)。

摘　　要：目的 了解大肠埃希菌和弗格森埃希菌dndC基因的携带情况,以及该基因与脉冲场凝胶电泳(PFGE)基因组降解表型之间的关系,探讨PFGE中菌株基因组降解的原因及处理方法。方法 使用PFGE对450株大肠埃希菌和弗格森埃希菌进行分析,设计引物检测dndC基因的携带情况并进行序列测定。通过dndC基因序列比对和进化树的构建,获得dndC基因型别以及其在不同型别大肠埃希菌中的分布。通过添加硫脲和消毒电泳仪研究基因组降解的处理方法。结果 450株大肠埃希菌和弗格森埃希菌中,降解的菌株为40株(8.89%)。dndC基因阳性菌株33株(7.33%),均出现了降解。大肠埃希菌携带的dndC基因可分为8个群。不同型别大肠埃希菌中均存在dndC基因阳性菌株,且阳性率存在差异。添加硫脲可以有效缓解PFGE分析中dndC基因导致的降解,电泳仪消毒可以缓解全胶降解的情况。结论 dndC基因会导致大肠埃希菌和弗格森埃希菌PFGE分析中基因组降解,添加硫脲可以缓解此降解。Objective To understand the carriage of dndC gene in Escherichia coli and E. fergusonii strains and the relationship between the dndC gene and the DNA degeneration phenotype, and explore the causes of DNA degeneration in pulsed-field gel electrophoresis(PFGE) and treatment methods. Methods A total of 450 E. coli and E. fergusonii strains were analyzed by PFGE. The primers were designed to detect and sequence the dndC gene. The phylogenetic tree was constructed based on the dndC genes obtained from this study and NCBI database to analyze the genotype of dndC gene and the distribution of dndC gene in different types of E. coli. Addition of thiourea and disinfection of electrophoresis equipment were used to study the treatment methods of DNA degeneration. Results There were 40 strains(8.89%) with the DNA degeneration phenotype by PFGE analysis and 33 strains of them were positive for dndC gene, accounting for 7.33% of 450E. coli and E. fergusonii strains. The dndC genes of E. coli were divided into eight groups. There were positive strains carrying dndC gene in all eight types of E. coli strains, and the positive rates of dndC gene in eight types of E. coli strains were different. Addition of thiourea and disinfection of electrophoresis equipment could effectively improve the DNA degeneration phenotype. Conclusion The dndC gene can cause DNA degradation of E. coli and E. fergusonii in PFGE analysis. Addition of thiourea was an effective method to ease the DNA degradation caused by dndC.

关 键 词：dndC基因 大肠埃希菌 弗格森埃希菌 脉冲场凝胶电泳 基因组降解 硫脲 

分 类 号：R211[医药卫生—中医学] R378