Ounissi H, Derlot E, Carlier C, Courvalin P
Unité des Agents Antibactériens, Unité Associée Centre National de la Recherche Scientifique 271, Institut Pasteur, Paris, France.
Antimicrob Agents Chemother. 1990 Nov;34(11):2164-8. doi: 10.1128/AAC.34.11.2164.
Aminoglycoside-resistant strains of Staphylococcus and Enterococcus, approximately 500 of each, were screened by dot blot hybridization for the presence of genes encoding aminoglycoside-modifying enzymes. The MICs of various aminoglycosides for the strains were determined, and the enzyme contents of the cells were inferred from the resistance phenotypes. The agreements (in percent) of the hybridization results with the deduced enzyme contents for Staphylococcus and Enterococcus species were, respectively, 80 and 87.6 for ANT(6) (aminoglycoside nucleotidyltransferase), 99.8 and 100 for both APH(3') (aminoglycoside phosphotransferase) and APH(2")-AAC(6') (aminoglycoside acetyltransferase), and 100 and 100 for ANT(4'). The weak correlation obtained with the probe for ANT(6) was due to the fact that gram-positive cocci can also be streptomycin resistant by synthesis of APH(3") or ANT(3")(9) and by ribosomal mutation. The remaining probes appeared to be specific: they hybridized with all the resistant clinical isolates but not with the susceptible strains. These results indicate that, except for streptomycin, nucleic acid hybridization is a valid approach for the detection and characterization of aminoglycoside resistance in gram-positive cocci.
对大约各500株的耐氨基糖苷类葡萄球菌和肠球菌菌株进行斑点印迹杂交,以检测编码氨基糖苷类修饰酶的基因的存在情况。测定了这些菌株对各种氨基糖苷类药物的最低抑菌浓度(MIC),并根据耐药表型推断细胞中的酶含量。葡萄球菌和肠球菌属杂交结果与推断的酶含量的一致性(百分比),对于ANT(6)(氨基糖苷类核苷酸转移酶)分别为80%和87.6%,对于APH(3')(氨基糖苷类磷酸转移酶)和APH(2")-AAC(6')(氨基糖苷类乙酰转移酶)均为99.8%和100%,对于ANT(4')为100%和100%。与ANT(6)探针获得的弱相关性是由于革兰氏阳性球菌也可通过合成APH(3")或ANT(3")(9)以及核糖体突变而对链霉素耐药。其余探针似乎具有特异性:它们与所有耐药临床分离株杂交,但不与敏感菌株杂交。这些结果表明,除链霉素外,核酸杂交是检测和鉴定革兰氏阳性球菌中氨基糖苷类耐药性的有效方法。