肺炎克雷伯菌的耐药性及其与生物膜形成的关系  

Drug Resistance of Klebsiella Pneumoniae and Its Relationship with Biofilm Formation

作　　者：陆颖 朱志军[1] 朱梅[1] LU Ying;ZHU Zhi-jun;ZHU Mei(Department of Laboratory Medicine,Chaohu Hospital,Anhui Medical University,Chaohu 238000,Anhui,China)

机构地区：[1]安徽医科大学附属巢湖医院检验科,安徽巢湖238000

出　　处：《医学信息》2021年第13期65-69,共5页Journal of Medical Information

基　　金：安徽医科大学校科研基金项目(编号:2020xkj216);安徽省卫生健康委科研项目(编号:AHWJ2021a013);安徽省教学研究项目(编号:2019jyxm1020)。

摘　　要：目的分析肺炎克雷伯菌的耐药机制及其与生物膜形成的关系。方法收集我院2018年8月-2019年6月临床分离的非重复肺炎克雷伯菌90株。采用微量肉汤稀释法检测抗菌药物的最低抑菌浓度(MIC),双纸片协同扩散法(DDST)及m CIM分别检测ESBLs和碳青霉烯酶,PCR检测耐药基因,通过96孔板结晶紫染色法测定菌株生物膜形成能力。结果 90株肺炎克雷伯菌中共检出66株可形成生物膜,占73.33%;生物膜阳性菌株对青霉素/β-内酰胺酶抑制剂、头孢菌素类、氨基糖苷类、复方新诺明和呋喃妥因类药物的耐药率均高于生物膜阴性菌株(P<0.05);MDR菌株中,生物膜阳性与生物膜阴性菌株的检出率分别为有65.15%和37.50%,生物膜阳性高于生物膜阴性菌株(P<0.02);37株DDST阳性菌株均携带ESBLs耐药基因,包括SHV型(97.30%),TEM型(62.16%),CTX-M-1组(54.05%)和CTX-M-9组(45.95%),且这些产ESBLs菌株中生物膜阳性与生物膜阴性菌株的检出率分别为48.48%和20.83%,差异有统计学意义(P<0.05);13株mCIM阳性菌株均携带KPC型,其中7株同时携带ESBLs耐药基因和KPC基因。产碳青霉烯酶的菌株中,生物膜阳性与阴性菌株相差不大。结论肺炎克雷伯菌形成生物膜可影响其对多种抗菌药物的耐药性,且发现了ESBLs耐药基因和KPC基因共存的菌株,须引起临床的重视,加强安全防护。Objective To analyze the resistance mechanism of Klebsiella pneumoniae and its correlation with biofilm formation.Methods A total of 90 non-repetitive Klebsiella pneumoniae strains isolated clinically in our hospital from August 2018 to June 2019 were collected.Micro broth dilution method was used to detect the minimum inhibitory concentration(MIC)of antibacterial drugs,the dual-disc cooperative diffusion method(DDST)and mCIM were used to detect ESBLs and carbapenemase respectively,and PCR was used to detect resistance genes,and crystallized through 96-well plates.Violet staining method was used to determine the biofilm forming ability of the strain.Results A total of 66 Klebsiella pneumoniae strains were found to form biofilms,accounting for 73.33%;The resistance rates of biofilm-positive strains to penicillin/β-lactamase inhibitors,cephalosporins,aminoglycosides,compound trimethoprim and nitrofurantoin drugs were higher than those of biofilm-negative strains(P<0.05);Among MDR strains,the detection rates of biofilm-positive and biofilm-negative strains were 65.15%and 37.50%,respectively,and biofilm-positive strains were higher than biofilm-negative strains(P<0.02);37 DDST-positive strains all carry ESBLs resistance genes,including SHV type(97.30%),TEM type(62.16%),CTX-M-1 group(54.05%)and CTX-M-9 group(45.95%)and the detection rates of biofilm-positive and biofilm-negative strains in these ESBLs-producing strains were 48.48%and 20.83%,respectively,the difference was statistically significant(P<0.05);The 13 mCIM-positive strains all carried the KPC type,and 7 of them carried both the ESBLs resistance gene and the KPC gene.Among the carbapenemase-producing strains,the biofilm-positive and negative strains are not much different.Conclusion The formation of biofilm by Klebsiella pneumoniae can affect its resistance to multiple antimicrobial drugs,and the discovery of strains with ESBLs resistance genes and KPC genes coexisting must arouse clinical attention and strengthen safety protection.

关 键 词：肺炎克雷伯菌 生物膜 抗菌药物 耐药基因 

分 类 号：R378.996[医药卫生—病原生物学]