耐碳青霉烯类铜绿假单胞菌耐药现状及整合子耐药基因分析

Analysis on current status of drug resistance of PA for antibiotics of carbapenems and resistance gene of integron

目的:研究深圳宝安区耐碳青霉烯类铜绿假单胞菌的耐药现状及其整合子耐药基因分布情况。方法:收集临床分离的耐碳青霉烯类铜绿假单胞菌93株,采用纸片扩散法(K-B法进行药敏检测、酶检测改良Hodge法检测碳青霉烯酶、B-last法检测超广谱β-内酰胺酶以及采用实时聚合酶链反应(PCR)对耐碳青霉烯类铜绿假单胞菌进行整合子耐药基因检测。结果:在93株耐碳青霉烯类铜绿假单胞菌中,对16种抗菌素耐药率>60%的占43.8%,其中复方新诺明和氨苄西林和(或)舒巴坦耐药率最高,占97.0%和95.0%,其次为亚胺培南和美洛培南,占20.0%和23.0%,多粘菌素E最低,占11.0%。在93株铜绿假单胞菌中碳青霉烯酶阳性率为80.7%,β-内酰胺酶阳性率为57.0%,改良Hodge法与PCR比较存在假阳性和假阴性,但差异无统计学意义;整合子耐药基因阳性率最高的为bla SHV-11,占81.7%;其次为bla KPC-2,占63.4%;bla SHV-142和bla IMP-4最低,占2.2%和4.3%;3株同时检出bla KPC-2和bla TEM-1基因,1株同时检出bla CTM-M-14和bla CTM-M-65基因,78株同时检出3种及以上耐药基因。结论:深圳宝安区铜绿假单胞菌对碳青霉烯类抗菌素耐药现象严重,最敏感为多粘菌素E,其次为亚胺培南和美洛培南;碳青霉烯酶和(或)β-内酰胺酶阳性是铜绿假单胞菌产生耐药的主要原因;整合子耐药基因普遍存在于铜绿假单胞菌,其中bla SHV-11和bla KPC-2基因为优势基因。

Objective:To understand current status of drug resistance of pseudomonas aeruginosa(PA)for antibiotics of carbapenems in Baoan district of Shenzhen and the distribution of resistance genes of their integron.Methods:93 strains of PA that were resistant for carbapenems were collected from clinical detection,and then K-B method was used to detect drug sensitivity,improved Hodge method was used to detect carbapenemases,and B-last method was used to detectβ-lactamase of super broad spectrum and real-time PCR was used to detect drug resistance gene of integron of PA for carbapenems.Results:In the 93 strains of PA,the drug resistance rate for 16 kinds of antibiotics was higher than 60%and these PA that were resistant for the 16 kinds of antibiotics were 43.8%of 93 strains.And in these antibiotics,the drug resistance rates of bactrim,ampicillin and(or)sulbactam were highest,and they were 97.0%and 95.0%,respectively.And the drug resistance rates of imipenem and meropenem were 20.0%and 23.0%,respectively,and the rate of polymyxin E was 11.0%.Besides,in the 93 strains of PA,the positive rates of carbapenemases andβ-lactamase were 80.7%and 57.0%,respectively.There were false positive and false negative existed in the comparison between improved Hodge method and PCR,while the difference was no significant.The positive rate of the drug resistance gene of integron of blaSHV-11 was highest(81.7%),and the rate of blaKPC-2 was second(63.4%),and the rates of blaSHV-142(2.2%)and blaIMP-4(4.3%)were lowest.There were 3 strains were simultaneously found out blaKPC-2 and blaTEM-1,and blaCTM-M-14 and blaCTM-M-65 were simultaneously found out in 1 strain.And in 78 strains,there were 3 and more than 3 kinds of drug-resistant genes were simultaneously found out.Conclusion:In Baoan district of Shenzhen,the phenomenon of drug resistance of PA for antibiotics of carbapenems was serious.And polymyxin E was the most sensitive,and the imipenem and meropenem were second.The positive of carbapenemases and(or)β-lactamase were the main reasons of drug resistance of PA.And the drug resistance genes of integron were ubiquity in PA,and in these genes,blaSHV-11 and blaKPC-2 were the protogene