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首页> 外文期刊>Journal of Microbiology, Biotechnology and Food Sciences >ANALYSES OF parC AND gyrA MUTATIONS IN CIPROFLOXACIN-RESISTANT AND SUSCEPTIBLE pseudomonas aeruginosa ISOLATED FROM SOIL BY PCR-RFLP AND SSCP METHOD
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ANALYSES OF parC AND gyrA MUTATIONS IN CIPROFLOXACIN-RESISTANT AND SUSCEPTIBLE pseudomonas aeruginosa ISOLATED FROM SOIL BY PCR-RFLP AND SSCP METHOD

机译：PCR-RFLP和SSCP方法分析耐环丙沙星和易感性铜绿假单胞菌中parC和gyrA突变

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The aims of this study were to assess the prevalence of gyrA and parC mutations in ciprofloxacin-resistant and susceptible Pseudomonas aeruginosa isolated from soil and to evaluate the suitability of the restriction fragment length polymorphism (RFLP) and single-strand conformation polymorphism (SSCP) techniques as screening methods for molecular characterizations of ciprofloxacin resistance. From the examined 21 P. aeruginosa isolates 9 strains were resistant to ciprofloxacin. These 21 P. aeruginosa isolates and one control strain were analyzed for alterations in the ciprofloxacin resistance determining region of gyrA and parC by RFLP and SSCP analyses. The PCR reaction confirmed the presence of the gyrA and parC genes in all of the strains. PCR-RFLP analyses with SacII for gyrA and HinfI for parC were performed as a screening method. We found that 18 and 17 out of 22 isolates have SacII and HinfI restriction site and 4 and 5 strains did not have the site recognized by these enzymes, respectively. Seven for gyrA and fourteen for parC PCR products were electrophoresed for SSCP. By SSCP analysis, 21 (in parC) and 15 (in gyrA) different band patterns were detected, and each pattern corresponded to a distinct mutation. As a result, the RFLP and SSCP methods are suitable for a molecular screening of ciprofloxacin resistant and susceptible P. aeruginosa isolates. SSCP analysis can also provide advantage for the detection of novel and multiple mutations. In addition, we can say that environmental monitoring followed by clinical surveillance can be successful in uncovering previously unrecognized cases of infection.
机译：这项研究的目的是评估从土壤中分离出的耐环丙沙星和敏感铜绿假单胞菌的gyrA和parC突变的发生率，并评估限制性片段长度多态性（RFLP）和单链构象多态性（SSCP）技术的适用性作为环丙沙星耐药分子表征的筛选方法。从所检查的21个铜绿假单胞菌分离物中，有9个菌株对环丙沙星具有抗性。通过RFLP和SSCP分析，分析了这21株铜绿假单胞菌分离株和1个对照菌株在gyrA和parC的环丙沙星抗性确定区域中的变化。 PCR反应证实了所有菌株中都存在gyrA和parC基因。作为筛选方法，使用SacII对gyrA和HinfI对parC进行PCR-RFLP分析。我们发现22个分离株中的18个和17个具有SacII和HinfI限制性酶切位点，而4个和5个菌株则没有被这些酶识别的位点。分别对gyrA的7个和parC PCR产物的14个进行SSCP电泳。通过SSCP分析，检测到21个（在parC中）和15个（在gyrA中）不同的条带模式，每种模式对应于一个独特的突变。结果，RFLP和SSCP方法适用于对环丙沙星耐药和易感的铜绿假单胞菌分离物进行分子筛选。 SSCP分析还可为检测新突变和多重突变提供优势。另外，我们可以说，先进行环境监测再进行临床监测，可以成功地发现以前无法识别的感染病例。

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