Luo Xia, Ye Chang-yun, Li Fang, Wang Hua, Ren Jun, Jing Huai-qi, Xu Jian-guo
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.
Zhonghua Liu Xing Bing Xue Za Zhi. 2007 Nov;28(11):1119-22.
To understand the variation of Shiga toxin (stx) genes of Escherichia coli O157:H7 strains isolated in China.
Polymerase chain reaction (PCR) was used to identity the types of stx genes and the nucleotide sequences of the amplified stex variants genes were determined. Compare to the cytotoxicity of Stx,variants were tested by HeLa cell assay.
We found novel stx2 genes in 3 of 289 strains of Shiga toxin-producing E. coli O157:H7 isolated from 1999 to 2002 in China. The novel stx2 genes were inserted by a 1.3-kb insertion sequence (IS) and the nucleotide sequences of IS showed 100% homology with that of IS1203 variant (IS1203v). The IS1203v inserted in the stx2 genes of three E. coli O157:H7 strains at different sites and the direction of the open reading frames (ORFs) of IS1203v of each strain was different. In addition to the above mentioned findings, the nucleotide sequences of three stx2 genes were completely identical and the type of the three Stx2 was Stx2 prototype. Compare to the cytotoxicity of Stx2 prototype, the novel Stx2 was found to be obviously lower.
E. coli O157:H7 strains harboring stx2::IS1203v genes were isolated in China. Consequently, the results of HeLa cell assay showed that the insertion of IS1203v could lead to low cytotoxicity of Stx2.
了解我国分离的大肠杆菌O157:H7菌株中志贺毒素(stx)基因的变异情况。
采用聚合酶链反应(PCR)鉴定stx基因类型,并测定扩增的stx变异基因的核苷酸序列。通过HeLa细胞试验检测与Stx细胞毒性相比,变异体的细胞毒性。
在1999年至2002年我国分离的289株产志贺毒素大肠杆菌O157:H7中,有3株发现了新的stx2基因。新的stx2基因被一个1.3 kb的插入序列(IS)插入,该IS的核苷酸序列与IS1203变异体(IS1203v)的核苷酸序列显示100%同源性。IS1203v插入到3株大肠杆菌O157:H7菌株的stx2基因的不同位点,且各菌株IS1203v的开放阅读框(ORF)方向不同。除上述发现外,3个stx2基因的核苷酸序列完全相同,3种Stx2的类型均为Stx2原型。与Stx2原型的细胞毒性相比,发现新的Stx2明显较低。
我国分离到携带stx2::IS1203v基因的大肠杆菌O157:H7菌株。因此,HeLa细胞试验结果表明,IS1203v的插入可导致Stx2的细胞毒性降低。