Vanhoof R, Content J, Van Bossuyt E, Nulens E, Sonck P, Depuydt F, Hubrechts J M, Maes P, Hannecart-Pokorni E
Pasteur Institute of Brabant, Brussels, Belgium.
J Antimicrob Chemother. 1993 Jul;32(1):23-35. doi: 10.1093/jac/32.1.23.
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.
利用聚合酶链反应（PCR）鉴定了编码氨基糖苷6'-N-乙酰基转移酶的基因。根据aacA基因的核苷酸序列，构建了四组引物，分别用于扩增209 bp、250 bp、260 bp和347 bp的DNA片段，这些片段分别对应四种已知的aacA基因，并在这些片段内设计了探针。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。这些引物与其相应的aacA基因发生反应，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检测法对161株表现出AAC(6')I活性的耐氨基糖苷临床分离株进行了检测。Tran Van Nhieu和Collatz（1987年）描述的基因是最常鉴定出的aacA基因。一株弗氏柠檬酸杆菌含有两个不同的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，未检测到任何已知aacA基因的特异性扩增DNA片段。
