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首页> 外文期刊>Journal of Clinical Microbiology >Rapid and specific detection of verotoxin genes in Escherichia coli by the polymerase chain reaction.
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Rapid and specific detection of verotoxin genes in Escherichia coli by the polymerase chain reaction.

机译：通过聚合酶链反应可快速，特异性地检测大肠杆菌中的维毒素基因。

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A set of four synthetic oligonucleotide probes derived from sequences of the VT1 (Shiga-like toxin I [SLT-I]) and VT2 (SLT-II) genes were used in a polymerase chain reaction (PCR) amplification procedure to detect these genes in some enteric pathogens. A total of 40 verotoxin-producing Escherichia coli strains and 43 isolates of other recognized enteric pathogens were studied. PCR amplification products identifying the VT1 and VT2 gene sequences were observed only in nucleic acid extracted from strains found to be VT positive in traditional tissue culture assays. Template nucleic acid extracted from other gram-negative bacteria was found to be negative with the exception of five isolates of Shigella dysenteriae type 1 in which good amplification with the VT1 probe was observed. The oligonucleotide probes clearly distinguished VT1 and VT2 strains of E. coli and did not give specific amplification with nucleic acid from VTe (a SLT-II variant)-producing E. coli. VT1 or VT2 genes or both were not detected in E. coli K-12 strain C600 or HB101 or in strains known to express other virulence factors, such as enterotoxins, adhesins, hemolysins, or unrelated cytotoxins. The sensitivity of the PCR procedure for detection of both VT1 and VT2 genes was determined to be 1 ng of total nucleic acid. Furthermore, the VT1 gene was easily detected when only 100 pg of nucleic acid was used as the template in the PCR procedure.
机译：在聚合酶链反应（PCR）扩增程序中使用了一组四个源自VT1（志贺样毒素I [SLT-I]）和VT2（SLT-II）基因序列的合成寡核苷酸探针，以检测这些基因。一些肠道病原体。总共研究了40种产生维毒素的大肠杆菌菌株和43种其他公认的肠病原体分离株。仅在从传统组织培养试验中发现为VT阳性的菌株中提取的核酸中观察到鉴定VT1和VT2基因序列的PCR扩增产物。发现从其他革兰氏阴性细菌中提取的模板核酸为阴性，除了五株痢疾志贺氏菌的五个分离株，其中观察到用VT1探针可进行良好扩增。寡核苷酸探针清楚地区分了大肠杆菌的VT1和VT2菌株，并且没有用产生VTe（一种SLT-II变体）的大肠杆菌的核酸进行特异性扩增。在大肠杆菌K-12菌株C600或HB101或已知表达其他毒力因子（例如肠毒素，黏附素，溶血素或无关细胞毒素）的菌株中未检测到VT1或VT2基因，或两者均未检测到。确定同时检测VT1和VT2基因的PCR程序的灵敏度为1 ng总核酸。此外，当在PCR程序中仅使用100 pg的核酸作为模板时，很容易检测到VT1基因。

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