Noppe-Leclercq I, Wallet F, Haentjens S, Courcol R, Simonet M
Laboratoire de bactériologie-hygiène, Centre hopitalier régional et universitaire, Lille, France.
Res Microbiol. 1999 Jun;150(5):317-22. doi: 10.1016/s0923-2508(99)80057-6.
Aminoglycoside resistance is common among strains of Acinetobacter baumannii responsible for nosocomial infections, and inactivation of these antibiotics by enzymatic modification is the main mechanism. Different types of aminoglycoside acetyltransferases (AAC), nucleotidyltransferases (ANT), and phosphotransferases (APH) are synthesized by clinical isolates, and several enzymes can be produced by a single strain. Using a multiplex PCR procedure carried out on bacterial thermolysates, we analyzed the aminoglycoside resistance gene content of strains belonging to eight clusters identified by pulsed-field gel electrophoresis. In a single reaction were combined three primer pairs in order to amplify the genes coding for AAC(6')-Ih, AAC(3)-I, and AAC(3)-II, three primer pairs for the genes coding for ANT(2'')-I, APH(3')-VI, and rRNA 16S as internal control, and finally two primer pairs for the genes coding for AAC(6')-Ib and APH(3')-I. According to the aminoglycoside resistance gene patterns, the strains of the eight clusters were distributed into seven classes. This simple and rapid (< 8 h) fingerprinting technique could be a useful tool for the epidemiological investigation of A. baumannii nosocomial infections.
氨基糖苷类耐药性在引起医院感染的鲍曼不动杆菌菌株中很常见，而通过酶修饰使这些抗生素失活是主要机制。临床分离株可合成不同类型的氨基糖苷类乙酰转移酶（AAC）、核苷酸转移酶（ANT）和磷酸转移酶（APH），并且单个菌株可产生几种酶。我们使用对细菌热裂解物进行的多重PCR程序，分析了通过脉冲场凝胶电泳鉴定的属于八个簇的菌株的氨基糖苷类耐药基因含量。在单个反应中组合了三对引物，以扩增编码AAC(6')-Ih、AAC(3)-I和AAC(3)-II的基因；三对引物用于扩增编码ANT(2'')-I、APH(3')-VI的基因以及作为内部对照的16S rRNA；最后两对引物用于扩增编码AAC(6')-Ib和APH(3')-I的基因。根据氨基糖苷类耐药基因模式，八个簇的菌株被分为七类。这种简单快速（<8小时）的指纹技术可能是鲍曼不动杆菌医院感染流行病学调查的有用工具。
