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用于检测和区分含有A、B和C型肠毒素基因以及中毒性休克综合征毒素1基因的金黄色葡萄球菌菌株的寡核苷酸探针。

Oligonucleotide probes for detection and differentiation of Staphylococcus aureus strains containing genes for enterotoxins A, B, and C and toxic shock syndrome toxin 1.

作者信息

Neill R J, Fanning G R, Delahoz F, Wolff R, Gemski P

机构信息

Department of Molecular Pathology, Walter Reed Army Institute of Research, Washington, D.C. 20307.

出版信息

J Clin Microbiol. 1990 Jul;28(7):1514-8. doi: 10.1128/jcm.28.7.1514-1518.1990.

Abstract

Different synthetic DNA nucleotide sequences were evaluated as gene probes for the specific detection and differentiation of Staphylococcus aureus strains encoding enterotoxins A (SEA), B (SEB), and C (SEC) and toxic shock syndrome toxin 1 (TSST-1). Identification of sequences unique to each toxin, based on knowledge of their nucleotide sequences, led to preparation of the specific 18-base oligonucleotide probes EA1 (encoding amino acids 177 to 182 of SEA), EB2 (encoding amino acids 105 to 110 of SEB), EC5 (encoding amino acids 125 to 131 of SEC1), and TS1 (encoding amino acids 160 to 166 of TSST-1). In colony blot hybridization analyses, these probes hybridized specifically with DNA from strains that produced the respective toxin serotypes. An excellent (greater than or equal to 93%) correlation between hybridization results (genotype) and toxin protein detection by an enzyme-linked immunosorbent assay (phenotype) was observed in the characterization of both reference and clinical strains of S. aureus for SEA, SEB, and TSST-1. A lower correlation (64%) for SEC reflected a lack of sensitivity in detecting toxin production. Our findings demonstrate that molecular DNA hybridization with synthetic oligonucleotide probes provides another approach for establishing the toxigenicity of S. aureus.

摘要

对不同的合成DNA核苷酸序列进行了评估,以作为基因探针,用于特异性检测和区分编码A(SEA)、B(SEB)和C(SEC)型肠毒素以及毒性休克综合征毒素1(TSST-1)的金黄色葡萄球菌菌株。基于对每种毒素核苷酸序列的了解,鉴定出每种毒素特有的序列,从而制备了特异性的18碱基寡核苷酸探针EA1(编码SEA的第177至182位氨基酸)、EB2(编码SEB的第105至110位氨基酸)、EC5(编码SEC1的第125至131位氨基酸)和TS1(编码TSST-1的第160至166位氨基酸)。在菌落印迹杂交分析中,这些探针与产生相应毒素血清型的菌株的DNA特异性杂交。在对金黄色葡萄球菌参考菌株和临床菌株的SEA、SEB和TSST-1特性进行表征时,观察到杂交结果(基因型)与通过酶联免疫吸附测定法检测毒素蛋白(表型)之间具有极好的(大于或等于93%)相关性。对于SEC,较低的相关性(64%)反映出在检测毒素产生方面缺乏敏感性。我们的研究结果表明,与合成寡核苷酸探针进行分子DNA杂交为确定金黄色葡萄球菌的产毒性提供了另一种方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/900e/267980/c16b62804cfe/jcm00055-0036-a.jpg

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