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目的 对福氏 2a志贺菌肠毒素ShET1和肠毒素ShET2进行基因分型 ,提高福氏 2a志贺菌同源克隆鉴定系统的分析水平。方法 综合运用质粒DNA分析、16srRNA基因探针分析及随机PCR分析方法对 93株分离自不同地区 ,不同时间的福氏 2a志贺菌进行多生物学标志分型 ,同时引入志贺菌肠毒素ShET1/ShET2基因PCR分析法检测福氏 2a志贺菌。结果 质粒DNA分析、RAPD分析、16srRNA基因探针分析三种方法联合运用 ,可将 93株S flexneri 2a分成 19个克隆群 ,克隆分辨率为 2 0 4 3% (19/ 93) ,引入毒素ShET1/ShET2基因PCR分析 ,可新增加 8个克隆群 ,克隆分辨率为 2 9 0 3% (2 7/ 93) ,克隆分辨率提高 8 6 0 % (8/ 93)。 93株福氏 2a志贺菌按志贺菌肠毒素分为 4种基因型 ,即 12株ShET1(- ) /ShET2 (+) ,14株ShET1(+) /ShET2 (- ) ,5 9株ShET1(+) /ShET2 (+) ,8株ShET1(- ) /ShET2 (- )。结论 在应用多生物学标志进行福氏 2a志贺菌同源克隆鉴定系统研究时 ,肠毒素ShET1/ShET2基因PCR分析是不可或缺的分析指标。
OBJECTIVE: To genotype Shiga toxin 2a Shiga toxin (ShET1) and enterotoxin (ShET2), and to improve the analysis of homologous clone identification system of Shigella flexneri 2a. Methods Multiplex biomarkers of 93 strains of Shigella flexneri 2a isolated from different regions and at different times were identified by using plasmid DNA analysis, 16srRNA gene probe analysis and random PCR analysis. Simultaneous introduction of Shigella enterotoxin ShET1 / ShET2 gene PCR analysis of Shigella flexneri 2a. Results Ninety-nine S flexneri 2a strains could be grouped into 19 clones using plasmid DNA analysis, RAPD analysis and 16srRNA gene probe analysis. The resolution of the clone was 2043% (19/93), and the toxin ShET1 / ShET2 gene, 8 clones could be newly added with a resolution of 29 0 3% (2 7/93) and a resolution of 86 0% (8/93). Ninety-three strains of Shigella flexneri were classified into four genotypes according to Shigella enterotoxins: 12 ShET1 (-) / ShET2 (+), 14 ShET1 (+) / ShET2 (+) / ShET2 (+) and 8 strains of ShET1 (-) / ShET2 (-). Conclusion The PCR analysis of ShET1 / ShET2 gene of enterotoxin is an indispensable index for the identification of Shigella flexneri 2a by using multiple biological markers.