Vertiev Iu V, Liaĭman M E, Ugriumova G A, Sergeeva T I
Klin Lab Diagn. 2000 Aug(8):46-50.
Polymerase chain reaction (PCR) was used for detection of pathogenic Clostridium botulinum, Clostridium perfringens, Clostridium difficile, and Escherichia coli. With this aim in view, primers to botulinic toxins types A, B, C1, D, E, F, and G, perfringens enterotoxin, difficile toxin, and types 1 and 2 Shigella-like toxins were chosen and synthesized. Optimal amplification conditions were selected for each pair of primers, with DNA and the respective agent as the reaction mixture matrices. PCR was highly specific and sensitive in all cases. Its sensitivity was 10-100 cells/sample. Among the tested C. botulinum, C. perfringens, C. difficile, and E. coli strains, specific amplification products of expected size were observed only in the strains containing the respective toxin genes. These findings recommend the use of these methods in clinical microbiology. Strains containing type 2 Shigella-like toxin gene were detected among E. coli strains isolated from patients with the hemolytic uremic syndrome, which for the first time indicates that the problem with E. coli epidemic strain O157 is valid for Russia. As a result of our studies, test systems for detection of types A, B, C, D, E, F, and G C. botulinum strains, C. perfringens and C. difficile, and E. coli O157 strains are now available.
聚合酶链反应(PCR)用于检测致病性肉毒梭菌、产气荚膜梭菌、艰难梭菌和大肠杆菌。基于此目的,选择并合成了针对A、B、C1、D、E、F和G型肉毒毒素、产气荚膜梭菌肠毒素、艰难梭菌毒素以及1型和2型志贺样毒素的引物。针对每对引物选择了最佳扩增条件,以DNA和相应试剂作为反应混合物基质。PCR在所有情况下都具有高度特异性和敏感性。其灵敏度为10 - 100个细胞/样本。在测试的肉毒梭菌、产气荚膜梭菌、艰难梭菌和大肠杆菌菌株中,仅在含有相应毒素基因的菌株中观察到预期大小的特异性扩增产物。这些发现表明这些方法可用于临床微生物学。在从溶血尿毒综合征患者分离出的大肠杆菌菌株中检测到含有2型志贺样毒素基因的菌株,这首次表明大肠杆菌流行菌株O157的问题在俄罗斯是存在的。作为我们研究的结果,现在已有用于检测A、B、C、D、E、F和G型肉毒梭菌菌株、产气荚膜梭菌和艰难梭菌以及大肠杆菌O157菌株的检测系统。