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目的了解2013年中国21个省市自治区桶装水来源铜绿假单胞菌的鉴定正确率,对我国桶装水来源铜绿假单胞菌的耐药状况进行初步分析。方法采用基于eta2和opr I两对基因的PCR方法对桶装水来源铜绿假单胞菌进行快速复核,使用Vitek GN生化鉴定卡进行生化验证,评价分离菌株的复核正确率和PCR方法的准确度,采用微量肉汤稀释法,对8类12种抗生素的耐药性进行测定。结果两对引物均能对铜绿假单胞菌扩增出预期的目的条带,但eta2具有更高的特异性;抽样进行生化试验结果显示2013年各地上报并运送的菌株复核正确率为100%;两种基因的PCR方法准确度均在95%以上,但均存在较低比例假阴性;2013年中国21个省市自治区上报的531株铜绿假单胞菌耐药率为11.68%(62/531),主要耐受多粘菌素B(5.27%,28/531)、氨曲南(4.14%,22/531)和美罗培南(3.01%,16/531),替卡西林/克拉维酸和替卡西林的中介率分别为43.31%(230/531)和25.42%(135/531)。结论各地上报分离的铜绿假单胞菌的准确性较高,PCR方法可对桶装水来源铜绿假单胞菌进行快速筛选,结合生化鉴定方法可准确鉴定。桶装水来源铜绿假单胞菌的耐药处于较低水平,但替卡西林/克拉维酸和替卡西林某种程度上显示了一定的耐药趋势,需要定期监测,以阐明桶装水中的铜绿假单胞菌的耐药特征和耐药趋势。
Objective To understand the identification accuracy rate of bottled water source Pseudomonas aeruginosa from 21 provinces in China in 2013 and to analyze the drug resistance status of bottled water source Pseudomonas aeruginosa in China. Methods A pair of PCR primers based on two pairs of eta2 and opr I genes was used for rapid review of bottled water-derived Pseudomonas aeruginosa. The Vitek GN biochemical identification card was used for biochemical verification to evaluate the correctness of the isolates and the accuracy of the PCR method , The micro broth dilution method was used to determine the drug resistance of 8 classes of 12 antibiotics. Results Two pairs of primers could amplify the expected band of Pseudomonas aeruginosa, but eta2 had higher specificity. The biochemical tests of samples showed that the correct rate of the strains reported and delivered in 2013 was 100% ; PCR methods of both genes were more than 95% accuracy, but both had a lower false negative rate; in 531 strains of P. aeruginosa reported in 21 provinces in China in 2013, the resistance rate was 11.68% (62 / 531), aztreonam (4.14%, 22/531) and meropenem (3.01%, 16/531), ticarcillin / clavulanic acid and The ticarcillin mediation rates were 43.31% (230/531) and 25.42% (135/531), respectively. Conclusion The reported accuracy of Pseudomonas aeruginosa isolated from different places is high. PCR method can be used to screen Pseudomonas aeruginosa from bottled water rapidly, and can be accurately identified by biochemical identification method. Bottled water sources Pseudomonas aeruginosa resistance is low, but ticarcillin / clavulanic acid and ticarcillin to some extent showed some resistance trends, the need for regular monitoring to clarify the bottled water The resistance characteristics and drug resistance of Pseudomonas aeruginosa.