Khan S A, Nawaz M S, Khan A A, Cerniglia C E
Division of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, Arkansas 72079, USA.
J Clin Microbiol. 2000 May;38(5):1832-8. doi: 10.1128/JCM.38.5.1832-1838.2000.
The transfer of ermA and ermC genes, the two most common resistance determinants of erythromycin resistance, was studied with Luria-Bertani broth in the absence of additional Ca(2+) or Mg(2+) ions. Fifteen human and five poultry isolates of Staphylococcus aureus, which were resistant to erythromycin but carried different genetic markers for erythromycin resistance, were used for conjugation. Since both the donors (Amp(s)-Tet(r)) and recipients (Amp(r)-Tet(s)) were resistant to erythromycin, the transconjugants were initially picked up as ampicillin- and tetracycline-resistant colonies. The resistance transfer mechanisms of the chromosomally located erythromycin rRNA methylase gene ermA and the plasmid-borne ermC gene were monitored by a multiplex PCR and gene-specific internal probing assay. Four groups of transconjugants, based upon the transfer of the ermA and/or ermC gene, were distinguished from each other by the use of this method. Selective antibiotic screening revealed only one type of transconjugant that was resistant to ampicillin and tetracycline. A high frequency of transfer (4.5 x 10(-3)) was observed in all of the 23 transconjugants obtained, and the direction of tetracycline and erythromycin resistance marker transfer was determined to be from poultry to clinical isolates. The transfers of the ermA and ermC genes were via transposition and transformation, respectively.
在不添加额外Ca(2+)或Mg(2+)离子的情况下,使用Luria-Bertani肉汤研究了红霉素抗性的两个最常见抗性决定因素ermA和ermC基因的转移。使用15株对红霉素耐药但携带不同红霉素抗性遗传标记的人源金黄色葡萄球菌分离株和5株家禽源金黄色葡萄球菌分离株进行接合试验。由于供体(氨苄青霉素抗性-四环素抗性)和受体(氨苄青霉素抗性-四环素敏感)均对红霉素耐药,因此最初将接合子挑选为对氨苄青霉素和四环素耐药的菌落。通过多重PCR和基因特异性内部探针检测法监测染色体定位的红霉素rRNA甲基化酶基因ermA和质粒携带的ermC基因的抗性转移机制。通过使用该方法,根据ermA和/或ermC基因的转移情况区分出四组接合子。选择性抗生素筛选仅揭示了一种对氨苄青霉素和四环素耐药的接合子类型。在所获得的所有23个接合子中均观察到高转移频率(4.5×10(-³)),并且确定四环素和红霉素抗性标记的转移方向是从家禽分离株到家临床分离株。ermA和ermC基因的转移分别通过转座和转化进行。