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使用DNA探针和聚合酶链反应检测粘质沙雷氏菌及其他革兰氏阴性菌中aac(6')-Ic基因的分布。

The use of a DNA probe and PCR to examine the distribution of the aac(6')-Ic gene in Serratia marcescens and other gram-negative bacteria.

作者信息

Snelling A M, Hawkey P M, Heritage J, Downey P, Bennett P M, Holmes B

机构信息

Department of Microbiology, University of Leeds, UK.

出版信息

J Antimicrob Chemother. 1993 Jun;31(6):841-54. doi: 10.1093/jac/31.6.841.

Abstract

A nucleotide probe for the chromosomal aminoglycoside 6' acetyltransferase gene (aac(6')-Ic) of Serratia marcescens was used in non-radioactive dot-blot hybridization experiments with 186 strains belonging to ten different species of Serratia. The gene was only detected in strains of S. marcescens (all strains tested), and positive hybridization was seen irrespective of whether or not strains were kanamycin-resistant. An additional 180 strains belonging to 28 Gram-negative bacterial species other than in the genus Serratia did not hybridize with the probe. A rapid PCR test for the aac(6')-Ic gene was developed and used to confirm that the aac(6')-Ic gene is only found in S. marcescens. Southern hybridization analysis of S. marcescens chromosomal DNA demonstrated that the gene was usually located on a PvuII fragment with a putative isoleucine tRNA-2 gene, but polymorphisms with respect to the size of this fragment were observed.

摘要

利用一种针对粘质沙雷氏菌染色体氨基糖苷6'-乙酰转移酶基因(aac(6')-Ic)的核苷酸探针,对属于10种不同沙雷氏菌的186株菌株进行了非放射性斑点杂交实验。该基因仅在粘质沙雷氏菌菌株(所有测试菌株)中检测到,无论菌株是否对卡那霉素耐药,均可见阳性杂交信号。另外180株属于沙雷氏菌属以外28种革兰氏阴性细菌的菌株,未与该探针杂交。开发了一种针对aac(6')-Ic基因的快速PCR检测方法,并用于确认aac(6')-Ic基因仅在粘质沙雷氏菌中发现。粘质沙雷氏菌染色体DNA的Southern杂交分析表明,该基因通常位于一个含有假定异亮氨酸tRNA-2基因的PvuII片段上,但观察到该片段大小存在多态性。

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