Tenover F C, Gootz T D, Gordon K P, Tompkins L S, Young S A, Plorde J J
J Infect Dis. 1984 Nov;150(5):678-87. doi: 10.1093/infdis/150.5.678.
Analysis of aminoglycoside-resistant Enterobacteriaceae isolated from patients at the Seattle Veterans Administration Medical Center indicated that a single 68-kilobase R factor was responsible for the epidemic spread of low-level resistance to gentamicin, kanamycin, and tobramycin. An examination, by means of the phosphocellulose paper binding assay, of resistant strains carrying this R factor resulted in the identification of a 2"-O-adenyltransferase [ANT(2")]-modifying enzyme. This enzyme was later detected in strains containing 150-kilobase plasmids. For more convenient monitoring of the dissemination of the ANT(2") gene among clinical isolates at the medical center, a DNA probe was developed by cloning of the ANT(2") structural gene from the 68-kilobase factor into pBR322. A 310-base pair Ava I restriction fragment isolated from the interior of the cloned ANT(2") gene was radiolabeled and used in Southern hybridization gels as a probe for plasmids isolated from aminoglycoside-resistant organisms. The probe proved to be highly specific and was more sensitive than enzymologic techniques for detection of the ANT(2") gene in clinical isolates with complex aminoglycoside resistance phenotypes.
对从西雅图退伍军人管理局医疗中心患者中分离出的耐氨基糖苷类肠杆菌科细菌的分析表明，一个单一的68千碱基R因子是导致对庆大霉素、卡那霉素和妥布霉素低水平耐药性流行传播的原因。通过磷酸纤维素纸结合试验对携带该R因子的耐药菌株进行检测，鉴定出一种2”-O-腺苷转移酶[ANT(2”)]修饰酶。后来在含有150千碱基质粒的菌株中也检测到了这种酶。为了更方便地监测医疗中心临床分离株中ANT(2”)基因的传播情况，通过将68千碱基因子中的ANT(2”)结构基因克隆到pBR322中，开发了一种DNA探针。从克隆的ANT(2”)基因内部分离出的一个310碱基对的Ava I限制性片段经放射性标记后，在Southern杂交凝胶中用作从耐氨基糖苷类生物体中分离出的质粒的探针。该探针被证明具有高度特异性，并且在检测具有复杂氨基糖苷类耐药表型的临床分离株中的ANT(2”)基因时比酶学技术更敏感。
