Real-time PCR detection of Pseudomonas aeruginosa in clinical and municipal wastewater and genotyping of the ciprofloxacin-resistant isolates
Real-time PCR detection of Pseudomonas aeruginosa in clinical and municipal wastewater and genotyping of the ciprofloxacin-resistant isolates
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DOI：
10.1111/j.1574-6941.2006.00100.x
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发表时间：
2006-07-01
影响因子：
4.2
通讯作者：
Obst, Ursula
中科院分区：
文献类型：
--
作者：
Schwartz, Thomas;Volkmann, Holger;Obst, Ursula
实时定量铜绿假单胞菌进行了各种废水系统，包括临床，城市废水和流入的污水处理厂。在临床废水中检测到最高浓度的铜绿假单胞菌特异性靶标。确定了由抑制或交叉反应导致的检测系统的局限性。从临床和城市废水中特异性富集后分离出耐环丙沙星的铜绿假单胞菌菌株。在某些情况下，它们也是从废水处理厂的废水和下游的河水中培养出来的。通过细菌图谱检测，共有119株分离株的表型特征为环丙沙星耐药。随后，通过微型测序确定基因gyrA密码子位置83和87、gyrB密码子位置466和parC密码子位置87和91中的氟喹诺酮耐药介导突变。环丙沙星耐药主要与细菌促旋酶和拓扑异构酶II基因gyrA密码子83位突变和parC密码子87或91位突变有关。将所有环丙沙星耐药铜绿假单胞菌菌株与来自氟喹诺酮耐药铜绿假单胞菌感染的临床数据的基因型进行比较。结果与临床分析的数据一致，除了在环丙沙星耐药铜绿假单胞菌废水分离株中未发现gyrA 87和gyrB突变。
Real-time quantification of Pseudomonas aeruginosa was performed in various wastewater systems including clinical, municipal wastewaters and inflow from a wastewater treatment plant. The highest concentrations of P. aeruginosa-specific targets were detected in clinical wastewaters. Limitations of the detection system resulting from inhibition or cross-reaction were identified. Ciprofloxacin-resistant P. aeruginosa strains were isolated after specific enrichment from clinical and municipal wastewaters. In some cases they were also cultivated from effluent of a wastewater treatment plant, and from its downstream river water. A total of 119 isolates were phenotypically characterized as ciprofloxacin-resistant via antibiogram testing. Subsequently, the fluoroquinolone-resistance-mediating mutations in the genes gyrA codon positions 83 and 87, gyrB codon position 466 and parC codon positions 87 and 91 were determined by mini-sequencing. Ciprofloxacin resistance was mainly associated with mutations in gyrA codon position 83 and parC mutation in codon positions 87 or 91 of the bacterial gyrase and topoisomerase II genes. All ciprofloxacin-resistant P. aeruginosa strains were compared with genotypes from clinical data of fluoroquinolone-resistant P. aeruginosa infections. The results were in agreement with data from clinical analyses, with the exception that no gyrA 87 and no gyrB mutations were found in ciprofloxacin-resistant P. aeruginosa wastewater isolates.