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Title: [Investigation of acquired drug-resistant genes and strains relationship in Pseudomonas aeruginosa isolated from burn patients].
Author: Fan YF, Cui SY, Zhang C, Xu XM.
Journal: Zhonghua Shao Shang Za Zhi; 2018 Feb 20; 34(2):83-87. PubMed ID: 29973025.
Abstract:
Objective: To investigate the acquired drug-resistant genes and strains relationship in 40 strains of Pseudomonas aeruginosa isolated from burn patients. Methods: Forty strains of Pseudomonas aeruginosa isolated from burn patients hospitalized in our burn department from January 2014 to December 2015 were selected, with 20 strains from each year. Kirby-Bauer paper disk diffusion method was used to detect sensitivity of the isolated Pseudomonas aeruginosa to 9 kinds of antibiotics of cefotaxime, ceftazidime, cefepime, imipenem, meropenem, gentamicin, amikacin, ciprofloxacin, and levofloxacin. Polymerase chain reaction was applied to detect 9 kinds of acquired β-lactamase antibiotics-resistant genes, outer membrane porin protein oprD2 genes, 12 kinds of acquired aminoglycosides antibiotics-resistant genes, and 6 kinds of acquired disinfectant-resistant genes and genetic marker genes of mobile genetic elements. Among the above genes, positive expression genes were verified by DNA sequencing and comparison. Sequences of twenty-eight acquired drug-resistant genes of the above 40 Pseudomonas aeruginosa strains were analyzed by unweighted pair-group method with arithmetic means cluster analysis. Results: Forty strains of Pseudomonas aeruginosa were resistant to the above 9 kinds of antibiotics. Two kinds of acquired β-lactamase antibiotics-resistant genes of bla(TEM), bla(CARB), 5 kinds of acquired aminoglycosides antibiotics-resistant genes of aac(6')-Ⅰb, aac(6')-Ⅱ, ant(2″)-Ⅰ, ant(3″)-Ⅰ, and rmtB, and 3 kinds of acquired disinfectant-resistant genes and genetic marker genes of mobile genetic elements of qacE△1-sul1, merA, and intⅠ1were detected in 40 strains of Pseudomonas aeruginosa with oprD2 gene deficiency. Forty strains aggregated obviously, with a total of 7 gene modes and 3 clones. Drug-resistant gene sequences of strains of number 2 to 4, 6 to 9, 11, 14, and 17 to 39 were similar and with close relationship. Drug-resistant gene sequences of number 12 and 13 strains were similar and with close relationship. Drug-resistant sequences of number 10 and 16 strains were similar and with close relationship. Conclusions: Genes of bla(TEM), bla(CARB), aac(6')-Ⅰb, aac(6')-Ⅱ, ant(2″)-Ⅰ, rmtB, qacE△1-sul1, merA, and intⅠ1 were prevalent in these strains of Pseudomonas aeruginosa with oprD2 gene deficiency isolated from burn patients, which may play key roles in resistance of Pseudomonas aeruginosa to β-lactamase, aminoglycoside, and quinolone antibiotics, and the drug-resistant phenotypes were in good coincidence with genotypes. Pseudomonas aeruginosa strains isolated from burn patients were with similar acquired drug-resistant genes and close relationship. 目的： 调查分离自烧伤患者的40株铜绿假单胞菌获得性耐药基因和菌株间的亲缘性。 方法： 选择笔者单位烧伤科2014年1月—2015年12月自烧伤患者分离出的40株铜绿假单胞菌，每年20株。采用K－B纸片扩散法检测分离出的菌株对头孢噻肟、头孢他啶、头孢吡肟、亚胺培南、美罗培南、庆大霉素、阿米卡星、环丙沙星、左氧氟沙星等9种抗菌药物的敏感性。采用PCR法检测9种β内酰胺类抗菌药物获得性耐药基因、外膜孔蛋白oprD2基因、12种氨基糖苷类抗菌药物获得性耐药基因、6种消毒剂获得性耐药基因与可移动遗传元件遗传标记基因，选取阳性表达基因进行DNA测序及比对。对40株铜绿假单胞菌的28种获得性耐药基因序列进行非加权组平均法聚类分析。 结果： 40株铜绿假单胞菌对上述9种抗菌药物均耐药。40株铜绿假单胞菌共检出bla(TEM)、bla(CARB)2种β内酰胺类抗菌药物获得性耐药基因，aac(6′)－Ⅰb、aac(6′)－Ⅱ、ant(2″)－Ⅰ、ant(3″)－Ⅰ、rmtB 5种氨基糖苷类抗菌药物获得性耐药基因，qacE△1－sul1、merA、intⅠ1 3种消毒剂获得性耐药基因及可移动遗传元件遗传标记基因，并均存在外膜孔蛋白oprD2基因缺失。40株铜绿假单胞菌聚集特征明显，共有7种基因模式和3个克隆株。2～4、6～9、11、14、17～39号菌株携带的耐药基因序列相似，亲缘性高；12、13号菌株携带的耐药基因序列相似，亲缘性高；10、16号菌株携带的耐药基因序列相似，亲缘性高。 结论：bla(TEM)、bla(CARB)、aac(6′)－Ⅰb、aac(6′)－Ⅱ、ant(2″)－Ⅰ、rmtB、qacE△1－sul1、merA、intⅠ1在烧伤患者铜绿假单胞菌菌株中流行且均存在oprD2基因缺失，可能是导致铜绿假单胞菌对β内酰胺类、氨基糖苷类、喹诺酮类抗菌药物均耐药的重要原因，耐药表型与基因型相符率高。烧伤患者检出铜绿假单胞菌耐药基因相似，菌株间亲缘性高。.

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