Use of the polymerase chain reaction (PCR) for the detection of aacA genes encoding aminoglycoside-6'-N-acetyltransferases in reference strains and gram-negative clinical isolates from two Belgium hospitals.
Use of the polymerase chain reaction (PCR) for the detection of aacA genes encoding aminoglycoside-6'-N-acetyltransferases in reference strains and gram-negative clinical isolates from two Belgium hospitals.
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使用聚合酶链式反应 (PCR) 检测来自比利时两家医院的参考菌株和革兰氏阴性临床分离株中编码氨基糖苷-6-N-乙酰转移酶的 aacA 基因。
DOI：
10.1093/jac/32.1.23
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
E. Hannecart
中科院分区：
文献类型：
--
作者：
R. Vanhoof;J. Content;E. van Bossuyt;E. Nulens;P. Sonck;F. Depuydt;J. Hubrechts;P. Maes;E. Hannecart
使用聚合酶链式反应 (PCR) 鉴定编码氨基糖苷 6'-N-乙酰转移酶的基因。基于 aacA 基因的核苷酸序列构建了四组引物，描绘了 209 bp、250 bp、260 bp 和 347 bp 的 DNA 片段，对四种已知的 aacA 基因具有特异性，以及这些片段内的探针。使用编码各种氨基糖苷修饰酶的参考菌株评估引物的特异性。引物与其相应的 aacA 基因反应，并且不与编码其他氨基糖苷修饰酶的基因发生交叉反应。使用 PCR 测定法测试了 161 个显示 AAC(6')I 活性的氨基糖苷类耐药临床分离株。 Tran Van Nhieu & Collatz (1987) 描述的基因是最常识别的 aacA 基因。弗氏柠檬酸杆菌的一种菌株含有两个不同的 aacA 基因。然而，在 46% 的菌株中，大多数是沙雷氏菌属。和不动杆菌属。没有检测到任何已知 aacA 基因的特定扩增 DNA 片段。
Genes encoding aminoglycoside 6'-N-acetyltransferases, were identified using the polymerase chain reaction (PCR). Four sets of primers delineating DNA fragments of 209 bp, 250 bp, 260 bp and 347 bp, specific for the four known aacA genes, and probes within these fragments, were constructed based on the nucleotide sequences of the aacA genes. The specificity of the primers was evaluated using reference strains encoding various aminoglycoside-modifying enzymes. The primers reacted with their corresponding aacA genes and did not cross-react with genes coding for other aminoglycoside-modifying enzymes. One hundred and sixty-one aminoglycoside resistant clinical isolates showing AAC(6')I activity were tested using the PCR assays. The gene described by Tran Van Nhieu & Collatz (1987) was the most frequently identified aacA gene. One strain of Citrobacter freundii contained two distinct aacA genes. However, in 46% of the strains, the majority being Serratia spp. and Acinetobacter spp. none of the specific amplified DNA fragments for any of the known aacA genes could be detected.