临床来源nct-CTXΦ样霍乱弧菌基因的多样性分析  

The diversity of nctCTXΦ prophage genome from clinical O1 strains of Vibrio choierae

作　　者：濮小英[1] 潘劲草[1] 孟冬梅[1] 叶榕[1] 黄志成[1] 

机构地区：[1]杭州市疾病预防控制中心微生物检验科,310006

出　　处：《中华微生物学和免疫学杂志》2007年第2期186-191,共6页Chinese Journal of Microbiology and Immunology

摘　　要：目的 了解临床来源霍乱弧菌nct-CTXΦ（不含霍乱毒素基因的CTXΦ）样基因特征。方法 对6株于2000-2004年自杭州散发腹泻患者分离的ctxA^-、tcpA^＋ 01群霍乱弧菌核糖体基因分型，PCR检测zot、noe、cep基因及串联CTXΦ基因组拷贝问的相邻片段；以zot、ace、cep基因探针South-ernblot分别检测nct-CTXΦ样基因的PstⅠ、HindⅢ、MluⅠ、BglⅡ酶切及HindⅢ、MluⅠ双酶切的限制性酶切长度多态性，结合拷贝间相邻片段HindⅢ酶切位点分析，构建nct-CTXΦ样基因组物理图谱。结果 6株01群霍乱弧菌中，核糖体基因分型为2种型别；1株无CTXΦ或nct-CTXΦ样基因组，5株存在nct-CTXΦ样基因组，且此5株菌株中存在3种类型nct-CTXΦ样基因组组成，分别为单拷贝长约6kb串联双拷贝、单拷贝长约5．8kb的串联双拷贝和串联4拷贝（2个单拷贝长约6kb，另2个单拷贝长约5．8kb）。结论 nct-CTXΦ霍乱弧菌可能具有引起腹泻的能力；临床和环境来源的霍乱弧菌中存在着一类多样性的nct-CTXΦ样噬菌体家族。Objedive To study molecular characteristic of ctxAB negative CTXΦ （nct-CTXΦ） prophage genomes from clinical O1 strains of lrtbrio cholerae. Methods The ribotyping characters of 6 V. cholerae strains with ctxA^- and tcpA^＋ were analyzed from sporadic diardma patients in Hangzhou from 2000 to 2004. To detemfine the existence of nct-CTXΦ genomes, gene zot, ace, cep and the contiguous sequences between two copies of nct- CTXΦ genomes were examined by polymecase chain reaction analysis. RFLP analysis with single restriction enzyme of Pst Ⅰ , HindⅢ, Mlu Ⅰ , or Bgl Ⅱ and double enzymes of Hindm and Mlu Ⅰ were performed with Southern blot of digoxigenin-labeled probes targeting to gene zot, ace, cep, respectively. Results Two close ribotypes were found in the 6 strains, whose 5 strains had nct-CTXΦ genomes, but neither CTXΦ nor net-CTXΦ genome was found in another strain. Among the former 5 strains, there were 3 kinds of net-CTXΦ prophage genomes. In 2 strains, two copies of nct-CTXΦ genome with single length of 6.0 kb were tandemly linked, in which there was not HindⅢ site located between the two genomes. In one strain, there were also two tandemly linked copies of nct-CTXΦ genome with single length of 5.8 kb without HindⅢ site between the two genoraes. In the other 2 strains, four tandemly linked copies of nct-CTXΦ genome were identified, in which two linked copies were 5.8 kb and the other two linked copies were 6.0 kb with an Hind m site between the two genomes. Conclusion V. cho/erae strains with ctxAB- and tcpA ＋ might be susceptible to sporadic diarrhea. There is one family of nct-CTXΦ pmphage genomes with diversity from clinical strains of V. cholerae.

关 键 词：霍乱弧菌 霍乱毒素 CTXΦ基因组 

分 类 号：R686[医药卫生—骨科学]