Eraç B, Gill A, Amyes S G, Gülay Z
Dokuz Eylül Universitesi Tip Fakültesi, Mikrobiyoloji ve Klinik Mikrobiyoloji Anabilim Dali, Izmir.
Mikrobiyol Bul. 2003 Apr-Jun;37(2-3):125-30.
In this study, the relationship between gyrA mutations and ciprofloxacin minimum inhibitory concentration (MIC) values was investigated in Escherichia coli strains. For this purpose, ciprofloxacin MIC values of 46 E. coli strains, isolated from out-patients and hospitalized patients, were determined by the agar dilution method. The "Quinolone Resistance Determining Region" (QRDR) of gyrA gene was amplified and restricted by Hinf-I enzyme. Ser-83 mutation was observed in all strains that have ciprofloxacin MIC values of 0.062 mg/L and higher. Afterwards, eight strains, that were found susceptible (MIC < 1 mg/L, n: 1), intermediate (MIC: 1-4 mg/L, n: 1) and high level resistant (MIC > 4 mg/L, n: 6) to ciprofloxacin, were chosen and mutations in QRDRs of these strains investigated by DNA sequence analysis. Ser 83 Leu mutation was found in all the chosen strains and Asp 87 Tyr or Asp 87 Asn mutations were also observed except the ciprofloxacin susceptible (MIC: 0.062 mg/L) one. In addition, base substitutions that don't lead to aminoacid changes were detected. The strain in which only Ser 83 Leu mutation was observed, showed high level nalidixic acid resistance (MIC > 256 mg/L). This fact was in favour of that, one mutation is enough to develop high level resistance to nalidixic acid. It was concluded that high level ciprofloxacin resistance requires at least two mutations in the QRDR of gyrA gene.
在本研究中,对大肠杆菌菌株中环丙沙星最低抑菌浓度(MIC)值与gyrA基因突变之间的关系进行了调查。为此,采用琼脂稀释法测定了从门诊患者和住院患者中分离出的46株大肠杆菌的环丙沙星MIC值。对gyrA基因的“喹诺酮耐药决定区”(QRDR)进行扩增并用Hinf-I酶进行酶切。在所有环丙沙星MIC值为0.062 mg/L及以上的菌株中均观察到Ser-83突变。之后,选择了8株对环丙沙星敏感(MIC < 1 mg/L,n = 1)、中介(MIC:1 - 4 mg/L,n = 1)和高水平耐药(MIC > 4 mg/L,n = 6)的菌株,并通过DNA序列分析研究这些菌株QRDR中的突变。在所有选定的菌株中均发现了Ser 83 Leu突变,除环丙沙星敏感株(MIC:0.062 mg/L)外,还观察到Asp 87 Tyr或Asp 87 Asn突变。此外,还检测到未导致氨基酸变化的碱基替换。仅观察到Ser 83 Leu突变的菌株表现出高水平的萘啶酸耐药性(MIC > 256 mg/L)。这一事实支持了一个突变就足以产生对萘啶酸的高水平耐药性这一观点。得出的结论是,环丙沙星高水平耐药需要gyrA基因的QRDR中至少有两个突变。